Substituted 2-quinolyl-oxazoles useful as PDE4 inhibitors

ABSTRACT

The invention claims compounds of the formula 
                         
wherein
 
     
       
         
         
             
             
         
       
         
         
           
             is a 5-membered heteroaryl; 
             X is S or O; 
             R 1  is H, alkyl, cycloalkyl, cylcoalkylalkyl-, —CH 2 F, —CHF 2 , —CF 3 , —C(O)alkyl or —C(O)NR 18 R 19 ; 
             R 3  and R 4  H, alkyl, hydroxyalkyl or —C(O)Oalkyl; 
             R 5  and R 6  are H, alkyl, hydroxyalkyl, alkoxyalkyl, mercaptoalkyl, —CH 2 F, —CHF 2 , —CF 3 , —C(O)OH or —C(O)Oalkyl; 
             R 7  is H, alkyl, alkenyl, hydroxyalkyl, cycloalkyl, alkoxyalkyl, aminoalkyl, (R 17 -phenyl)alkyl or —CH 2 —C(O)—O-alkyl; and R 8  comprises alkyl, heteroaryl, phenyl or cycloalkyl, or heterocycloalkyl, all optionally substituted, or a cycloalkyl- or heterocycloalkyl-substituted amide; or 
             R 7  and R 8  and the nitrogen to which they are attached together form an optionally substituted ring; and the remaining variables are as defined in the specification. Also claimed are pharmaceutical compositions, the use of the compounds as PDE4 inhibitors, and combinations with other actives.

CROSS REFERENCE TO RELATED APPLICATION

This application claims the benefit of U.S. Provisional Application 60/572,266, filed May 18, 2004.

FIELD OF THE INVENTION

The present invention relates to substituted 2-quinolyl-oxazoles, thiazoles, imidazoles and pyrazoles, pharmaceutical compositions comprising them, and their use as PDE4 inhibitors for treating a variety of diseases such as allergic and inflammatory diseases, CNS diseases and diabetes. Combinations with other agents useful in the treatment of several diseases are also claimed.

BACKGROUND

Phosphodiesterases are known to regulate cyclic AMP, and phosphodiesterase 4 (PDE4) has been shown to be the predominant regulator of cyclic AMP in respiratory smooth muscle and inflammatory cells. Inhibitors of PDE4 are useful in treating a variety of diseases, including allergic and inflammatory diseases, diabetes, central nervous system diseases, pain, and viruses that produce TNF.

Amino-substituted quinolyl PDE4 inhibitors are disclosed in U.S. Pat. No. 5,804,588; sulfonamide-substituted quinolyl PDE4 inhibitors are disclosed in U.S. Pat. No. 5,834,485; and (benzo-fused)heteroaryl-substituted PDE4 inhibitors are disclosed in U.S. Pat. No. 6,069,151.

SUMMARY OF THE INVENTION

The present invention relates to a compound having the structural formula I

or a pharmaceutically acceptable salt or solvate thereof, wherein

R is H or alkyl;

X is O or S;

R¹ is H, alkyl, cycloalkyl, cycloalkyl(C₁-C₄)alkyl-, —CH₂F, —CHF₂, —CF₃, —C(O)alkyl or —C(O)NR¹⁸R¹⁹;

R³ and R⁴ are independently selected from the group consisting of H, alkyl, hydroxyalkyl and —C(O)Oalkyl;

R⁵ and R⁶ are independently selected from the group consisting of H, alkyl, hydroxyalkyl, alkoxyalkyl, mercaptoalkyl, —CH₂F, —CHF₂, —CF₃, —C(O)OH, —C(O)Oalkyl and —C(O)NR⁴³R⁴⁴;

t is 1 or 2;

R⁷ is H, alkyl, alkenyl, hydroxyalkyl, cycloalkyl, alkoxyalkyl, aminoalkyl, (R¹⁷-phenyl)alkyl or —CH₂—C(O)—O-alkyl;

R⁸ is H, alkyl, alkenyl, alkoxy, alkoxyalkyl, hydroxyalkyl, dihydroxyalkyl, alkyl-NR¹⁸R¹⁹, cyanoalkyl, haloalkyl, R²³-heteroaryl, R²³-heteroarylalkyl, R³⁶-heterocycloalkyl, (R³⁶-heterocycloalkyl)alkyl, R¹⁷-phenyl, (R¹⁷-phenyl)alkyl, R¹⁷-naphthyl, (R¹⁷-naphthyl)alkyl, R¹⁷-benzyloxy, -alkyl-C(O)—NR¹⁸R¹⁹, -alkyl-C(O)—N(R³⁰)—(R²³-heteroaryl), -alkyl-C(O)—(R¹⁷-phenyl), -alkyl-C(O)—(R³⁶-heterocycloalkyl); -alkyl-N(R³⁰)—C(O)Oalkyl, -alkyl-N(R³⁰)—C(O)—NR¹⁸R¹⁹, -alkyl-N(R³⁰)—C(O)alkyl, -alkyl-N(R³⁰)—C(O)-(fluoroalkyl), -alkyl-N(R³⁰)—C(O)—(R³⁹-cycloalkyl), -alkyl-N(R³⁰)—C(O)—(R¹⁷-phenyl), -alkyl-N(R³⁰)—C(O)—(R²³-heteroaryl), -alkyl-N(R³⁰)—C(O)-alkylene-(R²³-heteroaryl), -alkyl-NH—SO₂—NR¹⁸R¹⁹, -alkyl-N(R³⁰)—(R¹⁷-phenyl), -alkyl-N(R³⁰)—(R²³-heteroaryl), -alkyl-O—(R¹⁷-phenyl), -alkyl-O—(R²³-heteroaryl), -alkyl-N(R³⁰)—SO₂-alkyl, alkylthioalkyl-, alkyl-SO₂-alkyl-, (R³⁵-phenylalkyl)-S-alkyl-, (hydroxyalkyl)-S-alkyl-, (alkoxyalkyl)-S-alkyl-, -alkyl-CO₂-alkyl, R⁴⁵-hydroxyalkyl, dihydroxyalkyl substituted by R¹⁷-benzyloxy, dihydroxyalkyl substituted by R¹⁷-phenyl, alkoxyalkyl substituted by R¹⁷-phenyl, (R¹⁷-phenyl)alkyl substituted by —CO₂alkyl, (R¹⁷-phenyl)alkyl substituted by —C(O)N(R³⁰)₂, alkyl substituted by (R²³-heteroaryl) and —C(O)NR³⁷R³⁸, haloalkyl substituted by CO₂alkyl, R¹²-cycloalkyl, (R¹²-cycloalkyl)alkyl,

or R⁷ and R⁸ and the nitrogen to which they are attached together form a ring system selected from the group consisting of

comprises an R³⁵-substituted 5 or 6-membered heteroaryl group fused to the piperidinyl or pyrrolidinyl ring;

p is 0 or 1;

q is 0 or 1;

the dotted line represents an optional double bond;

R⁹ is H, halo, alkyl, cycloalkyl, —CH₂F, —CHF₂ or CF₃;

R¹⁰, R¹¹, and R¹³ are independently selected from the group consisting of H and halo;

R¹² is 1-3 substituents independently selected from the group consisting of H, alkyl, hydroxy, alkoxy, hydroxyalkyl, alkoxyalkyl, —C(O)Oalkyl, —(CH₂)_(n)—N(R³⁰)—C(O)-cycloalkyl, —(CH₂)_(n)—N(R³⁰)—C(O)alkyl, —(CH₂)_(n)—N(R³⁰)—C(O)Oalkyl, —(CH₂)_(n)—N(R³⁰)—(R²³-heteroaryl), —(CH₂)_(n)—N(R³⁰)—C(O)—NR¹⁸R¹⁹, —(CH₂)_(n)—C(O)—NR¹⁸R¹⁹, R¹⁷-phenyl, R³⁵-heteroarylalkyl, R³⁵-heteroaryloxy, —C(O)-heterocycloalkyl, —O—C(O)-heterocycloalkyl, —O—C(O)—NR¹⁸R¹⁹, —NH—SO₂-alkyl, —NH—C(═NH)NH₂, and

or two R¹² substituents on the same carbon form ═O, ═NOR³⁰ or ═CH₂;

R¹⁴ is 1 or 2 substituents independently selected from the group consisting of H, OH, halo, alkyl, alkoxy, hydroxyalkyl, alkoxyalkyl, —CF₃, CN, R¹⁷-phenyl, (R¹⁷-phenyl)alkyl, —NR¹⁸R¹⁹, alkyl-NR¹⁸R¹⁹, —(CH₂)_(n)—C(O)OH, —(CH₂)_(n)—C(O)Oalkyl, —(CH₂)_(n)—C(O)alkyl, —(CH₂)_(n)—C(O)(R³⁵-phenyl), —(CH₂)_(n)—C(O)(R²³-heteroaryl), —(CH₂)_(n)—C(O)NR¹⁸R¹⁹, —(CH₂)_(n)—C(O)N(R³⁰)—(CH₂)_(n)—(R²³-heteroaryl), —(CH₂)_(n)—N(R³⁰)—C(O)alkyl, —(CH₂)_(n)—N(R³⁰)—C(O)-(fluoroalkyl), —(CH₂)_(n)—N(R³⁰)—C(O)-(cycloalkyl), —(CH₂)_(n)—N(R³⁰)—C(O)(R³⁵-phenyl), —(CH₂)_(n)—N(R³⁰)—C(O)(R²³-heteroaryl), —(CH₂)_(n)—N(R³⁰)C(O)NR¹⁸R¹⁹, —(CH₂)_(n)—N(R³⁰)—C(O)Oalkyl, —(CH₂)_(n)—N(R³⁰)cycloalkyl, —(CH₂)_(n)—N(R³⁰)(R¹⁷-phenyl), —(CH₂)_(n)—N(R³⁰)(R²³-heteroaryl), —(CH₂)_(n)—N(R¹⁸)SO₂alkyl, —(CH₂)_(n)—N(R²⁰)SO₂—(R₁₇-phenyl), —(CH₂)_(n)—N(R³⁰)SO₂—CF₃, —CH₂S(O)₀₋₂(R³⁵-phenyl), —(CH₂)_(n)—OC(O)N(R³⁰)alkyl, R²³-heteroaryl, (R²³-heteroaryl)alkyl, (R²³-heteroaryl)oxy, (R²³-heteroaryl)amino, —CH(OH)—(R¹⁷-phenyl), —CH(OH)—(R²³-heteroaryl), —C(═NOR³⁰)—(R¹⁷-phenyl), —C(═NOR³⁰)—(R²³-heteroaryl), morpholinyl, thiomorpholinyl,

w is 0 or 1;

or two R¹⁴ substituents and the carbon to which they are both attached form —C(═NOR³⁰)— or —C(O)—;

each n is independently 0, 1, 2 or 3;

R¹⁵ is H, alkyl, cycloalkyl, (cycloalkyl)alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl, —C(O)Oalkyl, —C(O)O(R³⁰-cycloalkyl), -alkyl-C(O)O-alkyl, —C(O)O-alkylene-(R³⁵-phenyl), R¹⁷-phenyl, (R¹⁷-phenyl)alkyl, —CH—(R¹⁷-phenyl)₂, R²³-heteroaryl, —(CH₂), —C(O)NR¹⁸R¹⁹, —SO₂-alkyl, —SO₂-cycloalkyl, —SO₂—CF₃, —SO₂—(R³⁵-phenyl), —SO₂—NR¹⁸R¹⁹, —C(O)alkyl, —C(O)-(fluoroalkyl), —C(O)—C(CH₃)(CF₃)₂, —C(O)—(R¹⁷-phenyl), —C(O)—(R²³-heteroaryl), —C(O)-hydroxyalkyl, —C(O)-alkoxyalkyl, —C(O)—(R³⁹-cycloalkyl), —C(O)-alkylene-(R¹⁷-phenyl), —C(O)-alkylene-(R²³-heteroaryl), —C(O)-alkylene-S—C(O)alkyl, —C(═S)—(R¹⁷-phenyl), hydroxyalkyl substituted by R¹⁷-phenyl, hydroxyalkyl substituted by R²³-heteroaryl, alkoxyalkyl substituted by R¹⁷-phenyl, alkoxyalkyl substituted by R²³-heteroaryl,

wherein z is 0, 1 or 2;

R¹⁶ is 1 to 4 substituents independently selected from the group consisting of H, alkyl, R¹⁷-phenyl, (R¹⁷-phenyl)alkyl, (R²³-heteroaryl)alkyl, hydroxyalkyl, alkoxyalkyl and —C(O)Oalkyl, or two R¹⁶ groups and the carbon to which they are both attached form —C(O)—;

R¹⁷ is 1 to 3 substituents independently selected from the group consisting of H, halo, alkyl, cycloalkyl, —OH, hydroxyalkyl, alkoxy, alkoxyalkyl, —CN, —CF₃, —OCF₃, —OCHF₂, —OCH₂F, —C(O)OH, —C(O)Oalkyl, —C(O)O—(R³⁵-phenyl), —C(O)alkyl, —C(O)—(R³⁵-phenyl), —SOalkyl, —SO₂alkyl, —SO₂—CF₃, alkylthio, —NR⁴³R⁴⁴, -alkyl-NR⁴³R⁴⁴, R³⁵-phenyl, R³⁵-phenoxy, R³⁵-heteroaryl, R³⁵-heteroaryloxy, R³⁶-heterocycloalkyl, —C(O)—(R³⁶-heterocycloalkyl), hydroxyalkyl-NH—, —C(O)N(R³⁰)₂, —N(R⁴³)—(R³⁵-cycloalkyl) and —C(═NOR³⁰); or two R¹⁷ substituents on adjacent carbon atoms together form —O—CH₂—O—, —O—(CH₂)₂—O—, —(CH₂)₂—O— or —O—CH₂—O—CH₂—;

R¹⁸ and R¹⁹ are independently selected from the group consisting of H, alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl, R¹⁷-phenyl, (R¹⁷-phenyl)alkyl, naphthyl and cycloalkyl;

R²⁰ is H, alkyl, or cycloalkyl;

R²² is 1 to 4 substituents independently selected from the group consisting of H, alkyl, hydroxy, alkoxy, halo, —CF₃, —NH₂ and R³⁵-phenyl;

R²³ is 1 to 4 substituents independently selected from the group consisting of H, alkyl, hydroxy, alkoxy, halo, —CF₃, —NR¹⁸R¹⁹, —CN, —C(O)Oalkyl, —SO₂-alkyl, —NHSO₂-alkyl, R³⁵-phenyl, R³⁵-heteroaryl, morpholinyl, and —(CH₂)_(n)—C(O)—N(R³⁰)₂;

R²⁴ is H, OH or alkoxy; or when the optional double bond is present, R²⁴ and the adjacent carbon atom form the double bond;

R²⁵ is H or R³⁵-phenyl;

R²⁷ is 1 to 3 substituents independently selected from the group consisting of H, halo, OH, alkyl, alkoxy, hydroxyalkyl, alkoxyalkyl, haloalkyl, —CN, —C(O)OH, —C(O)Oalkyl, —C(O)N(R³⁰)(R¹⁸), —C(O)—(R³⁶-hetercycloalkyl), R¹⁷-phenyl, (R¹⁷-phenyl)-alkyl, R²³-heteroaryl, (R²³-heteroaryl)alkyl, (R²³-heteroaryl)oxy, (R²³-heteroaryl)amino NR¹⁸R¹⁹, NR¹⁸R¹⁹-alkyl, —(CH₂)_(n)—N(R³⁰)—C(O)alkyl, —(CH₂)_(n)—N(R³⁰)—C(O)-(fluoroalkyl), —(CH₂)_(n)—N(R³⁰)—C(O)alkoxyalkyl, —(CH₂)_(n)—N(R³⁰)—C(O)(cycloalkyl), —(CH₂)_(n)—N(R³⁰)—(R²³-heteroaryl), —(CH₂)_(n)—N(R³⁰)—C(O)—(R²³-heteroaryl), —(CH₂)_(n)—N(R³⁰)—C(O)O-alkyl, —(CH₂)_(n)—N(R³⁰)—C(O)O—(CF₃-alkyl), —(CH₂)_(n)—N(R³⁰)—C(O)O—(R³⁹-cycloalkyl), —(CH₂)_(n)—N(R³⁰)—C(O)O-alkylene-cycloalkyl, —(CH₂)_(n)—N(R³⁰)—C(O)—N(R³⁰)(R²⁰), —(CH₂)_(n)—N(R³⁰)—SO₂-alkyl, —(CH₂)—N(R³⁰)—SO₂—CF₃, —(CH₂)_(n)—N(R³⁰)—SO₂—N(R³⁰)₂ and

or two R²⁷ groups and the carbon to which they are both attached form —C(═NOR³⁰)— or —C(O)—;

R²⁸ is H, alkyl, R³⁵-benzyl or -alkyl-C(O)O-alkyl;

R²⁹ is alkyl, haloalkyl, —C(O)Oalkyl, —C(O)alkyl, —C(O)CF₃, —C(O)—(R¹²-cycloalkyl), —C(O)—(R¹⁷-phenyl), —C(O)—(R²³-heteroaryl), —C(O)—(R³⁶-hetercycloalkyl), —SO₂-alkyl, —SO₂—(R³⁵-phenyl), —C(O)NR¹⁸R¹⁹, R³⁵-phenyl, (R³⁵-phenyl)alkyl or R²³-heteroaryl;

R³⁰ is independently selected from the group consisting of H, alkyl, R³⁵-benzyl and R³⁵-phenyl;

R³¹ is H, alkyl, R³⁵-benzyl or phenoxyalkyl;

R³³ is H, OH or alkoxy;

R³⁴ is H, alkyl, hydroxyalkyl, alkoxyalkyl or —C(O)Oalkyl;

R³⁵ is 1 to 3 substituents independently selected from the group consisting of H, halo, alkyl, OH, —CF₃, alkoxy, —CO₂alkyl and —N(R⁴³)(R⁴⁴);

R³⁶ is 1 or 2 substituents independently selected from the group consisting of H, alkyl, R¹⁷-phenyl, —OH, hydroxyalkyl, alkoxyalkyl, —C(O)Oalkyl and —NR¹⁸R¹⁹; or two R³⁶ groups and the carbon to which they are both attached form —C(═NOR³⁰)— or —C(O)—;

R³⁷ and R³⁸ are independently selected from the group consisting of H and alkyl, or R³⁷ and R³⁸ together are —(CH₂)₃— or —(CH₂)₄—, and together with the nitrogen to which they are attached, form a ring;

R³⁹ is H, OH, alkyl, alkoxy, or CF₃;

R⁴⁰ is —OR³⁰ or —NHC(O)alkyl;

R⁴¹ is H or —SO₂alkyl;

R⁴² is —(CH₂)_(n)—(R³⁵-phenyl), —(CH₂)_(n)—(R²³-heteroaryl), —C(O)Oalkyl or —C(O)alkyl;

R⁴³ and R⁴⁴ are independently selected from the group consisting of H and alkyl; and

R⁴⁵ is 1 or 2 substituents independently selected from the group consisting of halo, alkoxyalkyl, —CO₂alkyl, R¹⁷-phenyl, R²³-heteroaryl and cycloalkyl.

This invention also provides a method of treating diseases mediated by PDE 4, including allergic and inflammatory diseases, CNS diseases, and diabetes comprising administering an effective amount of at least one compound of formula I to a patient in need of such treatment.

In particular, this invention also provides a method of treating a PDE4 mediated disease or condition selected from the group consisting of: pain (e.g., acute pain, acute inflammatory pain, chronic inflammatory pain, and neuropathic pain), acute inflammation, chronic inflammation, rheumatoid arthritis, psoriasis, atopic dermatitis, asthma, COPD, adult respiratory disease, arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, stroke, ischemia reperfusion injury, renal reperfusion injury, glomerulonephritis, Parkinson's disease, Alzheimer's disease, mild cognitive impairment (MCI), depression, anxiety, graft vs. host reaction (i.e., graft vs. host disease), allograft rejections (e.g., acute allograft rejection, and chronic allograft rejection), acute respiratory distress syndrome, delayed type hypersensitivity reaction, atherosclerosis, cerebral ischemia, osteoarthritis, multiple sclerosis, angiogenesis, osteoporosis, gingivitis, respiratory viruses, herpes viruses, hepatitis viruses, HIV, Kaposi's sarcoma associated virus (i.e., Kaposi's sarcoma), meningitis, cystic fibrosis, pre-term labor, cough, pruritis, multi-organ dysfunction, psoriatic arthritis, herpes, encephalitis, traumatic brain injury, CNS tumors, interstitial pneumonitis, hypersensitivity, crystal induced arthritis, acute pancreatitis, chronic pancreatitis, acute alcoholic hepatitis, necrotizing enterocolitis, chronic sinusitis, ocular inflammation, corneal neovascularization, polymyositis, acne, esophagitis, glossitis, airflow obstruction, airway hyperresponsiveness (i.e., airway hyperreactivity), bronchiectasis, bronchiolitis, bronchiolitis obliterans (i.e., bronchiolitis obliterans syndrome), chronic bronchitis, dyspnea, emphysema, hypercapnea, hyperinflation, hypoxemia, hyperoxia-induced inflammations, hypoxia, pulmonary fibrosis, pulmonary hypertension, peritonitis associated with continuous ambulatory peritoneal dialysis (CAPD), granulocytic ehrlichiosis, sarcoidosis, small airway disease, ventilation-perfusion mismatching, wheeze, colds, gout, alcoholic liver disease, lupus, periodontitis, cancer, transplant reperfusion injury, early transplantation rejection (e.g., acute allograft rejection), airway hyperreactivity, allergic contact dermatitis, allergic rhinitis, alopecia areata, autoimmune deafness (including, for example, Meniere's disease), autoimmune hemolytic syndromes, autoimmune hepatitis, autoimmune neuropathy, autoimmune ovarian failure, autoimmune orchitis, autoimmune thrombocytopenia, chronic inflammatory demyelinating polyneuropathy, cirrhosis, dermatomyositis, diabetes, drug-induced autoimmunity, endometriosis, fibrotic diseases, gastritis, Goodpasture's syndrome, Graves' disease, Gullain-Barre disease, Hashimoto's thyroiditis, hepatitis-associated autoimmunity, HIV-related autoimmune syndromes and hematologic disorders, hypophytis, interstitial cystitis, juvenile arthritis, Langerhans' cell histiocytitis, lichen planus, metal-induced autoimmunity, myocarditis (including viral myocarditis), myositis, neuropathies (including, for example, IgA neuropathy, membranous neuropathy and idiopathic neuropathy), nephritic syndrome, optic neuritis, pancreatitis, post-infectious autoimmunity, primary biliary cirrhosis, reactive arthritis, ankylosing spondylitis, Reiter's syndrome, reperfusion injury, scleritis, scleroderma, secondary hematologic manifestation of autoimmune diseases (such as anemias), silicone implant associated autoimmune disease, Sjogren's syndrome, systemic lupus erythematosus, transverse myelitis, tubulointerstitial nephritis, uveitis, and vitiglio in a patient in need of such treatment comprising administering to said patient an effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof.

Compounds of formula I are preferably useful in treating pain (e.g., acute pain, acute inflammatory pain, chronic inflammatory pain, and neuropathic pain), acute inflammation, chronic inflammation, rheumatoid arthritis, psoriasis, atopic dermatitis, asthma, COPD, arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, septic shock, endotoxic shock, gram negative sepsis, toxic shock syndrome, stroke, ischemia reperfusion injury, glomerulonephritis, Parkinson's disease, Alzheimer's disease, mild cognitive impairment, depression, anxiety, graft vs. host reaction (i.e., graft vs. host disease), allograft rejections (e.g., acute allograft rejection, and chronic allograft rejection), delayed type hypersensitivity reaction, osteoarthritis, multiple sclerosis, angiogenesis, osteoporosis, HIV, cough, psoriatic arthritis, CNS tumors, necrotizing enterocolitis, airflow obstruction, airway hyperresponsiveness (i.e., airway hyperreactivity), bronchiolitis, chronic bronchitis, emphysema, pulmonary fibrosis, pulmonary hypertension, small airway disease, wheeze, lupus, cancer, transplant reperfusion injury, early transplantation rejection (e.g., acute allograft rejection), airway hyperreactivity, allergic contact dermatitis, allergic rhinitis, diabetes, juvenile arthritis, reactive arthritis, ankylosing spondylitis, reperfusion injury, and systemic lupus erythematosus.

More preferably, compounds of formula I are useful for treating COPD, asthma, IBD, dermatitis, MS, arthritis, Parkinson's disease, Alzheimer's disease, mild cognitive impairment, depression and anxiety.

Preferred veterinary uses for compounds of formula I include the treatment of dermatitis in dogs and the treatment of recurrent airway disease in horses.

This invention also provides a method of treating a PDE4 mediated disease or condition in a patient in need of such treatment comprising administering to said patient at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, in combination with at least one other medicament (e.g., a drug, agent or therapeutic) selected from the group consisting of:

-   -   a) disease modifying antirheumatic drugs;     -   b) nonsteroidal anitinflammatory drugs;     -   c) COX-2 selective inhibitors;     -   d) COX-1 inhibitors;     -   e) immunosuppressives;     -   f) steroids;     -   g) biological response modifiers;     -   h) other anti-inflammatory agents or therapeutics useful for the         treatment of chemokine mediated diseases; and     -   i) other agents or therapeutics useful for the treatment of         depression, anxiety, Alzheimer's Disease or Parkinson's Disease.

This invention also provides a method of treating a pulmonary disease (e.g., COPD, asthma or cystic fibrosis) in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, in combination with at least one compound selected from the group consisting of: glucocorticoids, 5-lipoxygenase inhibitors, β-2 adrenoceptor agonists, muscarinic M1 antagonists, muscarinic M3 antagonists, muscarinic M2 agonists, NK3 antagonists, LTB4 antagonists, cysteinyl leukotriene antagonists, bronchodilators, PDE4 inhibitors, PDE inhibitors, elastase inhibitors, MMP inhibitors, phospholipase A2 inhibitors, phospholipase D inhibitors, histamine H1 antagonists, histamine H3 antagonists, dopamine agonists, adenosine A2 agonists, NK1 and NK2 antagonists, GABA-b agonists, nociceptin agonists, expectorants, mucolytic agents, decongestants, antioxidants, anti-IL-8 anti-bodies, anti-IL-5 antibodies, anti-IgE antibodies, anti-TNF antibodies, IL-10, adhesion molecule inhibitors, and growth hormones.

This invention also provides a method of treating multiple sclerosis in a patient in need of such treatment comprising administering to said patient, a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, in combination with at least one compound selected from the group consisting of glatiramer acetate, glucocorticoids, methotrexate, azothioprine, mitoxantrone, chemokine inhibitors, and CB2-selective agents.

This invention also provides a method of treating multiple sclerosis in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, in combination with at least one compound selected from the group consisting of: methotrexate, cyclosporin, leflunimide, sulfasalazine, β-methasone, β-interferon, glatiramer acetate, and prednisone.

This invention also provides a method of treating rheumatoid arthritis in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, in combination with at least one compound selected from the group consisting of COX-2 inhibitors, COX inhibitors, immunosuppressives (e.g., methotrexate, cyclosporin, leflunimide and sulfasalazine), steroids (e.g., betamethasone, cortisone and dexamethasone), anti-TNF-α compounds, MMP inhibitors, glucocorticoids, chemokine inhibitors, CB2-selective inhibitors, and other classes of compounds indicated for the treatment of rheumatoid arthritis.

This invention also provides a method of treating stroke and ischemia reperfusion injury in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, in combination with at least one compound selected from the group consisting of thrombolitics (e.g., tenecteplase, TPA, alteplase), antiplatelet agents (e.g., gpIIb/IIIa), antagonists (e.g., abciximab and eftiifbatide), anticoagulants (e.g., heparin), and other compounds indicated for the treatment of rheumatoid arthritis.

This invention also provides a method of treating stroke and ischemia reperfusion injury in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, in combination with at least one compound selected from the group consisting of tenecteplase, TPA, alteplase, abciximab, eftiifbatide, and heparin.

This invention also provides a method of treating psoriasis in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, in combination with at least one compound selected from the group consisting of immunosuppressives (e.g., methotrexate, cyclosporin, leflunimide and sulfasalazine), steroids (e.g., β-methasone) and anti-TNF-α compounds (e.g., etonercept and infliximab).

This invention also provides a method of treating COPD in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof.

This invention also provides a method of treating arthritis in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof.

This invention also provides a method of treating osteoarthritis in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof.

This invention also provides a method of treating acute pain, acute inflammatory pain, chronic inflammatory pain, or neuropathic pain in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof.

This invention also provides a method of treating pain in a patient in need of such treatment comprising administering to said patient a therapeutically effective amount of at least one compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, and administering a therapeutically effective amount of at least one medicament selected from the group consisting of: NSAIDs, COXIB inhibitors, anti-depressants, and anti-convulsants.

This invention also provides a pharmaceutical composition comprising at least one (e.g., 1-3, usually 1) compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier. Preferred are oral dosage forms and dosage forms suitable for inhalation.

This invention also provides a pharmaceutical composition comprising at least one (e.g., 1-3, usually 1) compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, and at least one (e.g., 1-3, usually 1) other agent, medicament, antibody and/or inhibitor disclosed above, and a pharmaceutically acceptable carrier.

DETAILED DESCRIPTION

Preferred compounds of formula I are those wherein the quinolyl portion has the structure

More preferred are compounds wherein R¹⁰, R¹¹ and R¹³ are each H. Also preferred are compounds wherein R¹ is H, alkyl, cycloalkyl or CF₃; more preferably, R¹ is alkyl, especially methyl. Also preferred are compounds wherein R⁹ is H, alkyl or —CF₃, more preferably —CF₃.

In compounds of formula I, X is preferably O.

In compounds of formula I,

is preferably oxazolyl, more preferably

In compounds of formula I, R³ is preferably H, alkyl, hydroxalkyl or —C(O)Oalkyl, and R⁴ is H or alkyl. More preferably, R³ and R⁴ are each independently H or alkyl.

In compounds of formula I, R⁵ is preferably H, and R⁶ is preferably H, alkyl or hydroxyalkyl. When R⁶ is alkyl, it is preferably methyl, ethyl or isopropyl, more preferably methyl; when it is hydroxyalkyl, it is preferably hydroxymethyl or hydroxyethyl (i.e., —(CH₂)₂OH or —CH(OH)CH₃). In a more preferred embodiment, R⁵ is H and R⁶ is H, methyl or hydroxymethyl. Preferably, t is 1. When t is 2, preferably both R⁵ substituents and one R⁶ substituent are H and one R⁶ substituent is H or methyl.

Preferably, R⁵ and R⁶ have the following stereochemistry (i.e., R⁶ is “S”):

When R⁷ and R⁸ do not form a ring, the following definitions are preferred.

R⁷ is preferably H, alkyl, cycloalkyl, hydroxyalkyl or alkoxyalkyl. More preferably, R⁷ is H, alkyl, hydroxyalkyl, especially wherein alkyl is methyl or ethyl, and hydroxyalkyl is hydroxyethyl. Especially preferred are compounds wherein R⁷ is H or alkyl, especially H, methyl or ethyl, with H being most preferred.

R⁸ is preferably R¹²-cycloalkyl, (R¹²-cycloalkyl)alkyl, R⁴⁵-hydroxyalkyl, R¹⁷-phenyl, (R¹⁷-phenyl)alkyl, R²³-heteroaryl, (R²³-heteroaryl)alkyl, -alkyl-N(R³⁰)—C(O)—NR¹⁸R¹⁹, -alkyl-N(R³⁰)—C(O)alkyl, -alkyl-N(R³⁰)—C(O)—(R¹⁷-phenyl), -alkyl-N(R³⁰)—C(O)—(R²³-heteroaryl), -alkyl-N(R³⁰)—(R²³-heteroaryl),

More preferably, R⁸ is R¹²-cycloalkyl, R⁴⁵-hydroxyalkyl, (R¹⁷-phenyl)alkyl, R²³-heteroaryl, (R²³-heteroaryl)alkyl, -alkyl-N(R³⁰)—(R²³-heteroaryl), -alkyl-N(R³⁰)—C(O)alkyl,

(especially where p is 0) or

(especially where p is 1). Especially preferred are compounds wherein R⁸ is R¹²-cycloalkyl, R⁴⁵-hydroxyalkyl, (R¹⁷-phenyl)alkyl, (R²³-heteroaryl)alkyl, -alkyl-N(R³⁰)—C(O)-alkyl, -alkyl-N(R³⁰)—(R²³-heteroaryl) or

When R⁸ comprises R¹²-cycloalkyl, R¹² is preferably OH, —(CH₂)_(n)—N(R³⁰)—C(O)-cycloalkyl or —(CH₂)_(n)—N(R³⁰)—(R²³-heteroaryl), especially OH. When R⁸ is

n is preferably 0 and R²⁹ is preferably heteroaryl, —C(O)alkyl or —C(O)cycloalkyl. When R⁸ is R⁴⁵-hydroxyalkyl, R⁴⁵ is preferably R¹⁷-phenyl.

R³⁰ is preferably H.

Preferred heteroaryl groups include pyrimidyl, benzothienyl, benzofuranyl, indolyl, pyridyl and pyrazinyl.

Especially preferred are compounds of formula I wherein R⁷ is H and R⁸ is (R¹⁷-phenyl)alkyl, (R²³-heteroaryl)alkyl, R⁴⁵-hydroxyalkyl, -alkyl-N(R³⁰)—(R²³-heteroaryl) or

R¹⁷ is preferably 1-3 substituents selected from the group consisting of halogen, OH, alkoxy and alkyl; R²³ is preferably 1 or 2 substituents independently selected from the group consisting of H, alkyl, alkoxy and halogen; R⁴⁵ is preferably R¹⁷-phenyl, wherein R¹⁷ is 1-3 substituents selected from the group consisting of halogen, OH, alkoxy and alkyl; heteroaryl is pyrimidyl, benzothienyl, benzofuranyl, indolyl, pyridyl or pyrazinyl, and R³⁰ is H.

Also preferred are compounds of formula I wherein R⁷ and R⁸ and the nitrogen to which they are attached form

In the preferred compounds where R⁷ and R⁸ form

the optional double bond preferably is not present (i.e., a single bond is present). R¹⁴ is preferably selected from H, OH, alkoxy, —(CH₂)_(n)—N(R³⁰)(R²³-heteroaryl), R²³-heteroaryl or (R²³-heteroaryl)-alkyl. In a further preferred embodiment, one R¹⁴ is OH and the other R¹⁴ is R²³-heteroaryl; in another embodiment, one R¹⁴ is H and the other is (R²³-heteroaryl)-alkyl or —(CH₂)_(n)—N(R³⁰)(R²³-heteroaryl) (especially wherein n is 1).

In the preferred compounds where R⁷ and R⁸ form

q is preferably 1. R²⁷ is preferably 1-3 substituents independently selected from the group consisting of H, OH, alkyl, alkoxy, alkoxyalkyl, R¹⁷-phenyl, —C(O)OH, —C(O)Oalkyl, R²³-heteroaryl, (R²³-heteroaryl)amino and —(CH₂)_(n)—N(R³⁰)—C(O)(cycloalkyl), wherein n is 0.

In the preferred compounds where R⁷ and R⁸ form

R¹⁵ is preferably alkyl, R¹⁷-phenyl, R²³-heteroaryl, —C(O)alkyl, —C(O)(fluoroalkyl), —C(O)—(R²³-heteroaryl), —C(O)-alkoxyalkyl, —C(O)—(R³⁸-cycloalkyl), —SO₂-alkyl, —SO₂—NR¹⁸R¹⁹ or

R¹⁶ is preferably H, alkyl, or two R¹⁶ groups and the carbon to which they are attached form —C(O)—.

In the preferred compounds where R⁷ and R⁸ form

preferably p is 0, R³⁴ is hydrogen, and R³⁵ is 1 or 2 substituents independently selected from H, OH, halo and alkyl.

In the preferred compounds where R⁷ and R⁸ form

preferably p is 0, ring B is a pyrazolyl or thiazolyl ring, and R³⁵ is 1 or 2 substituents independently selected from H and alkyl.

As used above, and throughout the specification, the following terms, unless otherwise indicated, shall be understood to have the following meanings:

“Patient” includes both humans and animals.

“Mammal” means humans and other mammalian animals.

“Alkyl” means an aliphatic hydrocarbon group which may be straight or branched and comprising about 1 to about 6 carbon atoms in the chain. Branched means that one or more lower alkyl groups such as methyl, ethyl or propyl, are attached to a linear alkyl chain. Non-limiting examples of suitable alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl and n-pentyl.

“Alkenyl” means an aliphatic hydrocarbon group containing at least one carbon-carbon double bond and which may be straight or branched and comprising about 2 to about 6 carbon atoms in the chain. Branched means that one or more lower alkyl groups such as methyl, ethyl or propyl, are attached to a linear alkenyl chain. Non-limiting examples of suitable alkenyl groups include ethenyl, propenyl, n-butenyl, 3-methylbut-2-enyl and n-pentenyl.

“Alkylene” means a difunctional group obtained by removal of a hydrogen atom from an alkyl group that is defined above. Non-limiting examples of alkylene include methylene (i.e., —CH₂—), ethylene (i.e., —CH₂—CH₂—) and branched chains such as —CH(CH₃)—CH₂—.

“Heteroaryl” means a single ring, bicyclic or benzofused heteroaromatic group of 5 to 10 atoms comprised of 2 to 9 carbon atoms and 1 to 4 heteroatoms independently selected from the group consisting of N, O and S, provided that the rings do not include adjacent oxygen and/or sulfur atoms. N-oxides of the ring nitrogens are also included. Examples of single-ring heteroaryl groups are pyridyl, oxazolyl, isoxazolyl, oxadiazolyl, furanyl, pyrrolyl, thienyl, imidazolyl, pyrazolyl, tetrazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyrazinyl, pyrimidyl, pyridazinyl and triazolyl. Examples of bicyclic heteroaryl groups are naphthyridyl (e.g., 1,5 or 1,7), imidazopyridyl, pyridopyrimidinyl and 7-azaindolyl. Examples of benzofused heteroaryl groups are indolyl, quinolyl, isoquinolyl, phthalazinyl, benzothienyl (i.e., thianaphthenyl), benzimidazolyl, benzofuranyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl and benzofurazanyl. All positional isomers are contemplated, e.g., 2-pyridyl, 3-pyridyl and 4-pyridyl. The term R²³-heteroaryl refers to such groups wherein substitutable ring carbon atoms have a substituent as defined above. When the heteroaryl group is a benzofused ring, the substituents can be attached to either or both the phenyl ring portion and the heteroaromatic ring portion, and the heteroaryl group can be attached to the rest of the molecule either through the phenyl ring portion or the heteroaromatic ring portion.

“Cycloalkyl” means a non-aromatic mono- or multicyclic ring system comprising about 3 to about 10 carbon atoms, preferably about 3 to about 6 carbon atoms. Non-limiting examples of suitable monocyclic cycloalkyls include cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl and the like. Non-limiting examples of suitable multicyclic cycloalkyls include 1-decalin, norbornyl, adamantyl and the like. Monocyclic rings are preferred.

“Halo” means fluoro, chloro, bromo, or iodo groups. Preferred are fluoro, chloro or bromo, and more preferred are fluoro and chloro.

“Haloalkyl” means an alkyl as defined above wherein one or more hydrogen atoms on the alkyl is replaced by a halo group defined above; in particular, fluoroalkyl refers to an alkyl chain substituted by one or more fluoro atoms.

“Aminoalkyl” means an alkyl as defined above wherein a hydrogen atom on the alkyl is replaced by an amino (i.e., —NH₂) group.

“Heterocycloalkyl” means a non-aromatic saturated monocyclic or multicyclic ring system comprising about 3 to about 10 ring atoms, preferably about 5 to about 10 ring atoms, in which one or more, preferably 1, 2, 3 or 4, of the atoms in the ring system is independently selected from an element other than carbon, for example nitrogen, oxygen or sulfur, alone or in combination. There are no adjacent oxygen and/or sulfur atoms present in the ring system. Preferred heterocycloalkyls contain 5 to 6 ring atoms. The prefix aza, oxa or thia before the heterocycloalkyl root name means that at least a nitrogen, oxygen or sulfur atom respectively is present as a ring atom. The nitrogen or sulfur atom of the heterocyclyl can be optionally oxidized to the corresponding N-oxide, S-oxide or S-dioxide. Non-limiting examples of suitable monocyclic heterocyclyl rings include piperidyl, pyrrolidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiazolidinyl, 1,3-dioxolanyl, 1,4-dioxanyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like. The heterocycloalkyl group can be attached to the parent moiety through a ring carbon or a ring nitrogen.

“(Heterocycloalkyl)alkyl” means a heterocycloalkyl-alkyl group in which the heterocycloalkyl and alkyl groups are as defined above. The bond to the parent is through the alkyl.

“(Heteroaryl)alkyl” means a heteroaryl-alkyl- group in which the heteroaryl and alkyl are as previously described. Non-limiting examples of suitable heteroarylalkyl groups include pyridylmethyl, 2-(furan-3-yl)ethyl and quinolin-3-ylmethyl. The bond to the parent moiety is through the alkyl.

“(Phenyl)alkyl and “(naphthyl)alkyl similarly mean phenyl-alkyl and naphthyl-alkyl groups wherein the bond to the parent moiety is through the alkyl.

“Hydroxyalkyl” means a HO-alkyl- group in which alkyl is as previously defined. Non-limiting examples of suitable hydroxyalkyl groups include hydroxymethyl and 2-hydroxyethyl. Similarly, “dihydroxyalkyl” refers to a straight or branched alkyl chain substituted by two hydroxy groups.

“Alkoxy” means an alkyl-O— group in which the alkyl group is as previously described. Non-limiting examples of suitable alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy and n-butoxy. The bond to the parent moiety is through the ether oxygen.

“Alkylthio” means an alkyl-S— group in which the alkyl group is as previously described. Non-limiting examples of suitable alkylthio groups include methylthio, ethylthio and isopropylthio. The bond to the parent moiety is through the sulfur.

“Heteroarylamino” means an heteroaryl-NH— group in which the heteroaryl group is as previously described. Non-limiting examples of suitable heteroarylamino groups include pyrimidinyl-amino and pyrazinyl-amino. The bond to the parent moiety is through the amino nitrogen.

“Heteroaryloxy” means an heteroaryl-O— group in which the heteroaryl group is as previously described. Non-limiting examples of suitable heteroaryloxy groups include pyrimidinyl-O— and pyrazinyl-O—. The bond to the parent moiety is through the ether oxygen.

The term “hydroxyalkyl substituted by CO₂alkyl” means an alkyl chain substituted by a hydroxy group and a CO₂alkyl group. Similarly, terms such as “hydroxyalkyl substituted by R¹⁷-phenyl” means an alkyl chain substituted by a hydroxy group and a R¹⁷-phenyl group; “hydroxyalkyl substituted by R¹⁷-phenyl and alkoxy” means an alkyl group substituted by a hydroxy group, a R¹⁷-phenyl, and an alkoxy group. In each of these substituents and other similar substituents listed in the definitions, the alkyl chains can be branched.

Examples of moieties formed when two adjacent R¹⁷ groups form a ring with the carbons on the phenyl ring to which they are attached are:

When R⁷ and R⁸ together form

the dotted line indicates an optional double bond as defined above. When the double bond is absent, i.e., when a single bond is present, the one or two R¹⁴ substituents can be attached to the same or different ring carbons. When the double bond is present, only one R¹⁴ substituent can be attached to a carbon that is part of the double bond.

When R⁷ and R⁸ together form

the dotted line indicates an optional double bond as defined above. When the double bond is absent, i.e., when a single bond is present, R²⁴ can be H, OH or alkoxy and R²⁵ can be H or R³⁵-phenyl, but when the double bond is present, R²⁴ forms the double bond with the adjacent carbon and R²⁵ is H or R³⁵-phenyl. That is, the moiety has the structural formula

When R⁷ and R⁸ together form

it means that an optionally substituted fused bicyclic ring is formed, wherein the

portion comprises an R³⁵-substituted 5 or 6-membered heteroaryl group fused to the piperidinyl ring. Examples are:

The term “optionally substituted” means optional substitution with the specified groups, radicals or moieties, in available position or positions.

With reference to the number of moieties (e.g., substituents, groups or rings) in a compound, unless otherwise defined, the phrases “one or more” and “at least one” mean that there can be as many moieties as chemically permitted, and the determination of the maximum number of such moieties is well within the knowledge of those skilled in the art.

As used herein, the term “composition” is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts.

The wavy line

as a bond generally indicates a mixture of, or either of, the possible isomers, e.g., containing (R)- and (S)-stereochemistry. For example,

Lines drawn into the ring systems, such as, for example:

indicate that the indicated line (bond) may be attached to any of the substitutable ring carbon atoms.

As well known in the art, a bond drawn from a particular atom wherein no moiety is depicted at the terminal end of the bond indicates a methyl group bound through that bond to the atom, unless stated otherwise. For example:

It should also be noted that any carbon or heteroatom with unsatisfied valences in the text, schemes, examples, structural formulae, and any Tables herein is assumed to have the hydrogen atom or atoms to satisfy the valences.

Prodrugs and solvates of the compounds of the invention are also contemplated herein. The term “prodrug”, as employed herein, denotes a compound that is a drug precursor which, upon administration to a subject, undergoes chemical conversion by metabolic or chemical processes to yield a compound of formula I or a salt and/or solvate thereof. A discussion of prodrugs is provided in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems (1987) Volume 14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in Drug Design, (1987) Edward B. Roche, ed., American Pharmaceutical Association and Pergamon Press, both of which are incorporated herein by reference thereto.

“Solvate” means a physical association of a compound of this invention with one or more solvent molecules. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In certain instances the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid. “Solvate” encompasses both solution-phase and isolatable solvates. Non-limiting examples of suitable solvates include ethanolates, methanolates, and the like. “Hydrate” is a solvate wherein the solvent molecule is H₂O.

“Effective amount” or “therapeutically effective amount” is meant to describe an amount of compound or a composition of the present invention effective in inhibiting PDE4 and thus producing the desired therapeutic effect in a suitable patient.

The compounds of formula I form salts which are also within the scope of this invention. Reference to a compound of formula I herein is understood to include reference to salts thereof, unless otherwise indicated. The term “salt(s)”, as employed herein, denotes acidic salts formed with inorganic and/or organic acids, as well as basic salts formed with inorganic and/or organic bases. In addition, when a compound of formula I contains both a basic moiety, such as, but not limited to a pyridine or imidazole, and an acidic moiety, such as, but not limited to a carboxylic acid, zwitterions (“inner salts”) may be formed and are included within the term “salt(s)” as used herein. Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, although other salts are also useful. Salts of the compounds of the formula I may be formed, for example, by reacting a compound of formula I with an amount of acid or base, such as an equivalent amount, in a medium such as one in which the salt precipitates or in an aqueous medium followed by lyophilization. Acids (and bases) which are generally considered suitable for the formation of pharmaceutically useful salts from basic (or acidic) pharmaceutical compounds are discussed, for example, by S. Berge et al, Journal of Pharmaceutical Sciences (1977) 66(1) 1-19; P. Gould, International J. of Pharmaceutics (1986) 33 201-217; Anderson et al, The Practice of Medicinal Chemistry (1996), Academic Press, New York; in The Orange Book (Food & Drug Administration, Washington, D.C. on their website); and P. Heinrich Stahl, Camille G. Wermuth (Eds.), Handbook of Pharmaceutical Salts: Properties, Selection, and Use, (2002) Int'l. Union of Pure and Applied Chemistry, pp. 330-331. These disclosures are incorporated herein by reference thereto.

Exemplary acid addition salts include acetates, adipates, alginates, ascorbates, aspartates, benzoates, benzenesulfonates, bisulfates, borates, butyrates, citrates, camphorates, camphorsulfonates, cyclopentanepropionates, digluconates, dodecylsulfates, ethanesulfonates, fumarates, glucoheptanoates, glycerophosphates, hemisulfates, heptanoates, hexanoates, hydrochlorides, hydrobromides, hydroiodides, 2-hydroxyethanesulfonates, lactates, maleates, methanesulfonates, methyl sulfates, 2-naphthalenesulfonates, nicotinates, nitrates, oxalates, pamoates, pectinates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propionates, salicylates, succinates, sulfates, sulfonates (such as those mentioned herein), tartarates, thiocyanates, toluenesulfonates (also known as tosylates,) undecanoates, and the like.

Exemplary basic salts include ammonium salts, alkali metal salts such as sodium, lithium, and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, aluminum salts, zinc salts, salts with organic bases (for example, organic amines) such as benzathines, diethylamine, dicyclohexylamines, hydrabamines (formed with N,N-bis(dehydroabietyl)ethylenediamine), N-methyl-D-glucamines, N-methyl-D-glucamides, t-butyl amines, piperazine, phenylcyclohexylamine, choline, tromethamine, and salts with amino acids such as arginine, lysine and the like. Basic nitrogen-containing groups may be quarternized with agents such as lower alkyl halides (e.g. methyl, ethyl, propyl, and butyl chlorides, bromides and iodides), dialkyl sulfates (e.g. dimethyl, diethyl, dibutyl, and diamyl sulfates), long chain halides (e.g. decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides), aralkyl halides (e.g. benzyl and phenethyl bromides), and others.

All such acid salts and base salts are intended to be pharmaceutically acceptable salts within the scope of the invention and all acid and base salts are considered equivalent to the free forms of the corresponding compounds for purposes of the invention.

Compounds of formula I, and salts, solvates and prodrugs thereof, may exist in their tautomeric form (for example, as an amide or imino ether). All such tautomeric forms are contemplated herein as part of the present invention.

All stereoisomers (for example, geometric isomers, optical isomers and the like) of the present compounds (including those of the salts, solvates and prodrugs of the compounds as well as the salts and solvates of the prodrugs), such as those which may exist due to asymmetric carbons on various substituents, including enantiomeric forms (which may exist even in the absence of asymmetric carbons), rotameric forms, atropisomers, and diastereomeric forms, are contemplated within the scope of this invention. Individual stereoisomers of the compounds of the invention may, for example, be substantially free of other isomers, or may be admixed, for example, as racemates or with all other, or other selected, stereoisomers. The chiral centers of the present invention can have the S or R configuration as defined by the IUPAC 1974 Recommendations. The use of the terms “salt”, “solvate” “prodrug” and the like, is intended to equally apply to the salt, solvate and prodrug of enantiomers, stereoisomers, rotamers, tautomers, racemates or prodrugs of the inventive compounds.

Polymorphic forms of the compounds of Formula I, and of the salts, solvates and prodrugs of the compounds of Formula I, are intended to be included in the present invention.

This invention also includes the compounds of this invention in isolated and pure form.

Compounds of formula I can be prepared by known methods from starting materials either known in the art or prepared by methods known in the art. Non-limiting examples of suitable methods are illustrated in the following schemes.

In the schemes, the quinolyl portion is shown as the preferred structure, but those skilled in the art will recognize that other substitutions on the quinolyl portion can be made by these procedures. Also, one skilled in the art will recognize that the schemes show the significant steps of the procedures, and that the synthesis of compounds of formula I may require the need for the protection of certain functional groups during the preparation of the compounds; the synthesis of compounds also may require the reduction of a reducible functional group or the oxidation of an oxidizable functional group.

Step a

Formation of the oxazole ring can be accomplished by a number of methods including, but not limited to the following.

Using the appropriate starting materials, both the amine and ester functional groups can be incorporated when the oxazole ring is synthesized.

Step b

Introduction of the ester moiety COOR² can be accomplished stepwise by reaction with phosphorous oxychloride and subsequent oxidation of the intermediate aldehyde to the carboxylic acid and further esterification. Alternatively, reaction at this position can proceed with a Lewis acid such as zinc triflate and an acid chloride.

Step c

Introduction of the R moiety can be accomplished by deprotonation with a strong base such as n-butyl lithium, sec-butyl lithium, lithium diisopropylamine or lithium hexamethyldisilazide, followed by addition of an aldehyde or alkyl halide. This reaction can use a variety of solvents including diethyl ether, THF, dioxane, hexane, toluene, HMPA, DMPU and TMEDA.

Step d

Activation of the R moiety in (3) can be accomplished by several different methods. If R is an alkyl moiety, halogenation, for example with bromine or N-bromo-succinimide and an initiator such as benzoyl peroxide, AIBN or light in carbon tetrachloride as the solvent provides (5) as a halide. If R incorporates an ester or alcohol functional group, through appropriate oxidation or reduction reactions, the aldehyde or ketone functional group can be obtained for further reaction in Step e through a reductive amination reaction. If R incorporates an ester, ketone, or aldehyde functional group, appropriate reduction reaction with a hydride such as NaBH₄, LiBH₄, LiAlH₄, or diisobutylaluminum hydride will provide the alcohol moiety. This alcohol can be activated by conversion, for example, to the corresponding mesylate, tosylate, chloride, bromide or iodide.

Step e

Introduction of the amine moiety in (6) can be accomplished by an alkylation reaction on (5) if X is a leaving group such as chloride, bromide, mesylate or tosylate. This reaction can use a variety of bases including TEA, DIPEA, N-methyl morpholine, pyridine, dimethylaminopyridine, imidazole, K₂CO₃, Cs₂CO₃, potassium t-butoxide, and NaOH, and can be done in a variety of solvents including DMF, dimethylacetamide, THF, dioxane, CH₃CN, toluene, CH₂Cl₂ and dichloroethane. Alternatively, if the —C(X)(R⁵)(R⁶) moiety incorporates a ketone or aldehyde functional group, the amine moiety can be introduced through a reductive amination reaction. Suitable reducing reagents for this reaction include NaBH₃CN, sodium triacetoxyborohydride in a mixture of solvents including THF, dioxane, CH₃CN, toluene, CH₂Cl₂, dichloroethane, methanol, ethanol, trifluoroethanol. The reductive amination reaction may require the addition of a drying agent such as sieves or MgSO₄, or azeotropic removal of water or the addition of a Lewis acid such as titanium isopropoxide. In addition, the ketone or aldehyde moiety can be converted into an oxime with hydroxylamine and a variety of bases such as pyridine, TEA, sodium acetate, and Na₂CO₃. The oxime can be reduced to an amine.

Step f

Hydrolysis of ester (6) to acid (7) can be accomplished with a suitable base such as NaOH, LiOH, sodium methoxide, sodium ethoxide, K₂CO₃, Cs₂CO₃, BCl₃, potassium t-butoxide, TEA, DBU and DIPEA in a mixture of solvents including water, methanol, ethanol, isopropanol, CH₂Cl₂, THF, diethyl ether and dioxane.

Step g

Amide bond formation to obtain (8) can be accomplished by formation of the acid chloride, a mixed anhydride, or activated ester and addition of the appropriate amine. A variety of suitable amide bond coupling reagents such as HATU, CDI, EDC, DCC, PyBOP, polymer supported CDI, polymer supported EDC and the like, with or without HOBt, can be used. These coupling reagents can be used with a suitable base such as TEA, DIPEA, N-methyl morpholine, pyridine, dimethylaminopyridine, DBU, imidazole and the like in a mixture of solvents including DMF, dimethylacetamide, THF, dioxane, CH₃CN, N-methylpyrrolidine, CH₂Cl₂, and dichloroethane.

Abbreviations used in the above general schemes and in the following examples, as well as throughout the specification, are as follows: Me (methyl); Bu (butyl); Et (ethyl); Ac (acetyl); Boc or BOC (t-butoxycarbonyl); DMF (dimethyl-formamide); THF (tetrahydrofuran); DIPEA (diisopropylethylamine); RT (room temperature); HOBt (hydroxybenzotriazole); TFA (trifluoroacetic acid); TEA (triethyl amine); KHMDS (potassium bis(trimethylsilyl)amide); TLC (thin layer chromatography); EDC (1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride); HMPA (hexamethylphosphoramide); DMPU (1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone); TMEDA (N,N,N′,N′-tetramethyletheylenediamine); HATU (O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyl uranium hexafluoro-phosphate); NBS (N-bromosuccinimide); DCC (1,3-dicyclohexylcarbodiimide); DEC (1,2-diethylaminoethyl chloride hydrochloride); TMSCN (trimethylsilylcyanide); CDI (carbonyldiimidazole); PyBOP (benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate).

EXAMPLE 1

Step 1: SOCl₂ (26.7 ml, 367 mmol) was added to a mixture of compound 1 (40 g, 147 mmol) in dry toluene (300 ml) and DMF (0.4 ml). The mixture was heated at 70° C. for 2 h, then the excess of SOCl₂ and solvents were evaporated to dryness to obtain compound 2 as an off white-solid (41 g).

Step 2: A solution of compound 2 (41 g, 141 mmol) in CH₂Cl₂ (200 ml) was added slowly to a solution of L-threonine methyl ester HCl salt (29 g, 170 mmol) in CH₂Cl₂ (200 ml) and DIPEA (38 g, 296 mmol) at 0° C. The solution was stirred at 0° C., then warmed to RT over 3 h. After 3 h at RT, the mixture was washed with aqueous NH₄Cl solution, then the solid was precipitated in the organic layer and filtered off to give compound 3 (54 g) as a white solid. MS: C₁₇H₁₇F₃N₂O₅ [M+1]⁺387.1.

Step 3: SOCl₂ (76.8 ml, 645 mmol) was added through a syringe to a suspension of compound 3 (50 g, 129 mmol) in dry CH₂Cl₂ (500 ml) cooled to −45 0° C. The mixture was stirred at −45° C. for 1 h, then warmed up to RT slowly. After the reaction was complete, solvent and excess SOCl₂ were evaporated. The residue was dissolved in CH₂Cl₂ (800 ml) and washed with saturated NaHCO₃ solution (3×600 ml), dried (Na₂SO₄), filtered and concentrated to give compound 4 as a beige solid (43 g, 120 mmol, 93%). MS: C₁₇H₁₅F₃N₂O₄ [M+1]⁺369.1.

Step 4: DBU (13.9 ml, 93 mmol) was added via a syringe to a solution of compound 4 (31 g, 84 mmol) in dry CH₂Cl₂ (300 ml) at 0° C., followed by the addition of BrCCl₃ (9.1 ml, 93 mmol). The mixture was stirred at 0° C. for 2 h, then at RT overnight. The reaction was quenched with 0.15 N HCl (400 ml) and extracted with CH₂Cl₂ (2×100 ml). The organic layer was dried with Na₂SO₄, filtered and concentrated to give crude title compound 5 (35 g). The crude material was triturated with MeOH (200 ml) and 23.5 g of compound 5 was collected as a pale yellow solid. MS: C₁₇H₁₃F₃N₂O₄ [M+1]⁺367.1.

EXAMPLE 2

Step 1: NBS (23.5 g, 132 mmol) and benzoyl peroxide (1.4 g, 5.75 mmol) were added to a mixture of compound 5 (42 g, 115 mmol) in dry CCl₄ (550 ml). The mixture was refluxed for 3 h, then concentrated by evaporating off most of the solvent. Saturated NH₄Cl solution was added and the product was extracted from the aqueous layer with CH₂Cl₂ (2×300 ml). The organic fractions were combined, dried (Na₂SO₄), filtered and evaporated. The crude material was triturated with MeOH to give compound 6 as a white solid (49.5 g, 110 mmol, 96%). MS: C₁₇H₁₂F₃BrN₂O₄ [M+1]⁺Br^(79,81) 445.1, 447.1.

Step 2: Potassium phthalimide (20.6 g, 111 mmol) was added to a solution of compound 6 (49.5 g, 111 mmol) in dry DMF (650 ml) at RT. After stirring at RT for 2 h, the reaction mixture was poured into an ice water bath (1.5 L). The resultant yellow precipitate was collected, washed with water, and dried at 45° C. under vacuum to give compound 7 as a yellow solid (56 g, 110 mmol). MS: C₂₅H₁₆F₃N₃O₆ [M+1]⁺512.0.

Step 3: BCl₃ (1 M in CH₂Cl₂, 78 ml, 78 mmol) solution was added to a solution of compound 7 (10 g, 19.57 mmol) in dry CH₂Cl₂ (400 ml) at −15° C. After the addition of BCl₃, the mixture turned yellow and precipitate started to form. The reaction was warmed to 0° C. After the reaction was complete (checked by TLC), the mixture was poured into ice-water (600 ml). The yellow precipitate was filtered, washed with water, and dried (Na₂SO₄) to give the title compound 8 as a yellow solid (8.5 g, 17.1 mmol, 87%). MS: C₂₄H₁₄F₃N₃O₆ [M+1]⁺498.1.

EXAMPLE 3

Step 1: To a suspension of compound 8 (0.35 g, 0.7 mmol) in dry DMF (16 ml), compound 9 (3-aminomethyl benzothiophene) (0.11 g, 0.7 mmol), DI PEA (0.18 g, 1.4 mmol) and HATU (0.53 g, 1.4 mmol) were added at RT. After 30 ml, the reaction mixture was poured into cold water (30 ml). The precipitate was filtered, washed with water and dried under vacuum to give crude compound 10 (0.45 g, 0.7 mmol) as a yellow solid.

Step 2: The crude material of compound 10 (0.45 g, 0.7 mmol) was treated with absolute EtOH (15 ml) and 98% hydrazine (0.22 g, 7 mmol) at RT overnight. The reaction mixture was evaporated and purified on a Biotage (40 M) system, eluting with 3% NH₄OH:CH₃OH (1:9)/97% CH₂Cl₂. Compound 11 was obtained as a pure yellow solid (0.22 g, 0.43 mmol, 61% yield) which was converted to its HCl salt by treatment with 1.2 equivalent of 4 N HCl/dioxane in CH₂Cl₂. Compound 12 was obtained by evaporating off solvents and excess acid. MS (M+1): m/e. 513.

EXAMPLE 4

A series of aromatic or heteroaromatic amide analogs (compound 13) was made by methods analogous to those described for compound 12 in Example 3 or via an alternative coupling method by treatment of compound 8 (0.2 mmol) either with aromatic or heteroaromatic amine reagent (0.2 mmol), DEC (0.24 mmol), HOBT (0.24 mmol), and TEA (0.24 mmol) in DMF (2.5 ml) at RT overnight. Water (3 ml) was added to the reaction and precipitate was collected, rinsed with water, and vacuum dried at 40° C. The phthalamido protecting group of the coupled product was removed with 98% hydrazine in EtOH (as in step 2 of Example 3) and purified by silica gel chromatography [5% NH₄OH—CH₃OH (1:9) in 95% CH₂Cl₂] or by preparative Gilson Prep column (XTerra RP C₁₈, 5 μm) chromatography, gradient eluted with 0.5% TFA in (9:1) (H₂O—CH₃CN) to 0.5% TFA in CH₃CN:H₂O (8:2). Compound 13 was obtained as free form or as a TFA salt, depending on the method of purification. The free form of compound 13 was treated with 1.2 equivalent of HCl to give compound 13 as a HCl salt. The data for compound 13 analogs are listed as follows:

13

Cpd. MS No. Structure (M + 1) 13-1 

487   13-2 

471   13-3 

471   13-4 

475   13-5 

489   13-6 

487   13-7 

491   13-8 

525   13-9 

515   13-10 

515   13-11 

515   13-12 

501   13-13 

525   13-14 

475   13-15 

509   13-16 

525   13-17 

491   13-18 

475   13-19 

491   13-20 

525   13-21 

509   13-22 

493   13-23 

493   13-24 

493   13-25 

535   13-26 

487   13-27 

517   13-28 

509   13-29 

541   13-30 

523   13-31 

487   13-32 

505   13-33 

483   13-34 

539   13-35 

513   13-36 

469   13-37 

457   13-38 

487   13-39 

537   13-40 

517   13-41 

497   13-42 

525   13-43 

499   13-44 

547   13-45 

485   13-46 

483   13-47 

485   13-48 

501   13-49 

501   13-50 

485   13-51 

523   13-52 

526   13-53 

541   13-54 

525   13-55 

507   13-56 

535   13-57 

515   13-58 

499   13-59 

533   13-60 

503   13-61 

483   13-62 

473   13-63 

521   13-64 

521   13-65 

521   13-66 

507   13-67 

499   13-68 

499   13-69 

485   13-70 

485   13-71 

485   13-72 

485   13-73 

538   13-74 

500   13-75 

487   13-76 

565   13-77 

505   13-78 

505   13-79 

505   13-80 

523   13-81 

527   13-82 

513   13-83 

527   13-84 

527   13-85 

555   13-86 

555   13-87 

509   13-88 

515   13-89 

508   13-90 

482   13-91 

499   13-92 

473   13-93 

515   13-94 

503   13-95 

458   13-96 

488   13-97 

487   13-98 

499   13-99 

519   13-100

527   13-101

547   13-102

531   13-103

571   13-104

515   13-105

545   13-106

13-107

496   13-108

511   13-109

523   13-110

527   13-111

550   13-112

536   13-113

561   13-114

567   13-115

581   13-116

595   13-117

525   13-118

532   13-119

539   13-120

569   13-121

533   13-122

546   13-123

555   13-124

540   13-125

541   13-126

536   13-127

633   13-128

617   13-129

664   13-130

539   13-131

533   13-132

568   13-133

554   13-134

437   13-135

453   13-136

497   13-137

591   13-138

635   13-139

629   13-140

526   13-141

511   13-142

582   13-143

569   13-144

574   13-145

629   13-146

447   13-147

538   13-148

589   13-149

573   13-150

579   13-151

579   13-152

576   13-153

512   13-154

580   13-155

553   13-156

535   13-157

619   13-158

605   13-159

513   13-160

501   13-161

501   13-162

636   13-163

604   13-164

473.5 13-165

501   13-166

501   13-167

518   13-168

487   13-169

487   13-170

601   13-171

645   13-172

580   13-173

552   13-174

541   13-175

568   13-176

584   13-177

636   13-178

652   13-179

478   13-180

478   13-181

542   13-182

542   13-183

568   13-184

585   13-185

515   13-186

637   13-187

542   13-188

530   13-189

540   13-190

510   13-191

609   13-192

629   13-193

541   13-194

482   13-195

514   13-196

512   13-197

517   13-198

518   13-199

529   13-200

546   13-201

568   13-202

584   13-203

543   13-204

543   13-205

561   13-206

582   13-207

517   13-208

541   13-209

536   13-210

624   13-211

541   13-212

554   13-213

550   13-214

550   13-215

526   13-216

529   13-217

576   13-218

573   13-219

521   13-220

566   13-221

471   13-222

485   13-223

522   13-224

536   13-225

543   13-226

522   13-227

505   13-228

510   13-229

545   13-230

467   13-231

519   13-232

536   13-233

453   13-234

514   13-235

534   13-236

590   13-237

513   13-238

520   13-239

467   13-240

581   13-241

581   13-242

544   13-243

529   13-244

477   13-245

529   13-246

520   13-247

534   13-248

569   13-249

542   13-250

507   13-251

625   13-252

577   13-253

521   13-254

522   13-255

604   13-256

593   13-257

556   13-258

575   13-259

602   13-260

465   13-261

590   13-262

590   13-263

594   13-264

593   13-265

542   13-266

560   13-267

587   13-268

527   13-269

526   13-270

541   13-271

541   13-272

599   13-273

602   13-274

520   13-275

527   13-276

580   13-277

529   13-278

528   13-279

529   13-280

542   13-281

500   13-282

505   13-283

501   13-284

536   13-285

557   13-286

558   13-287

559   13-288

524   13-289

544   13-290

541   13-291

616   13-292

464   13-293

518   13-294

472   13-295

502   13-296

502   13-297

507   13-298

586   13-299

516   13-300

502   13-301

516   13-302

497   13-303

517   13-304

527   13-305

527   13-306

498   13-307

522   13-308

535   13-309

556   13-310

EXAMPLE 5

Step 1: Glycine ethyl ester hydrochloride (14 g, 100 mmol) was mixed with TEA (29 ml, 200 mmol) in dry CH₂Cl₂ and cooled in an ice-water bath. Compound 2 (80 mmol) (see Example 1) in dry CH₂Cl₂ (150 ml) was transferred by cannulation into the above cooled solution. The resulting mixture was allowed to warm to RT slowly. After 2 h, reaction was complete and water (300 ml) was added to dissolve the TEA salt. The organic layer was separated, washed with 5% HCl solution, then water, dried (Na₂SO₄), filtered and concentrated to give a crude solid, Compound 14, which was used in the next step without further purification.

Step 2: Compound 14 (7.2 g, 20 mmol) was mixed with Lawesson's reagent (5.5 g, 13.6 mmol) in anhydrous THF (100 ml) and heated to 78° C. for 40 min. After cooling to RT, THF was removed and product was purified by silica chromatography, eluting with 100% CH₂Cl₂ to 5% EtOAc in CH₂Cl₂, to give compound 15 as a yellow product.

Step 3: Compound 15 (3.9 g, 10 mmol) was dissolved in dry CH₂Cl₂ (40 ml) and cooled to −78° C. Trimethyloxonium tetrafluoroborate (1.6 g, 11 mmol) was added in one portion. The resulting mixture was then stirred in an ice-water bath for 2 h. NaHCO₃ solution was added to quench the reaction. The organic layer was separated, washed with H₂O, dried (Na₂SO₄), and evaporated to give compound 16 as a crude solid which was used in the next reaction without purification.

Step 4a:

A neat liquid of cyanuric fluoride (3.4 ml, 40 mmol) was added dropwise to a cooled solution of N-[(1,1-dimethylethoxy)carbonyl]-L-alanine (compound 17) (3.90 g, 20 mmol) in pyridine (1.78 ml, 22 mmol) and dry CH₂Cl₂ (50 ml) at −40° C. The reaction was kept at −30° C. to −10° C. for 2 h. After 2 h, crushed ice and CH₂Cl₂ (100 ml) were added. After stirring for 5 min, the mixture was filtered twice, first with a coarse, then with a medium glass filter funnel. The clear solution was separated and the organic phase was washed with H₂O, dried (Na₂SO₄), filtered and concentrated at RT to give compound 18 as a white solid (3.59 g, 18.7 mmol) with a 94% yield.

Step 5: Compound 18 (2.87 g, 15 mmol) was added to a solution of compound 16 (5.0 g, 12.5 mmol) in dry THF (60 ml). The reaction mixture was cooled to −78° C. and KHMDS (0.5 M in toluene) (52.5 ml, 26.25 mmol) was added dropwise over 40 min. During the addition of the first equivalent of base, the reaction mixture turned a deep blue color, which disappeared immediately. A deep brown color was formed when the second equivalent of base was added. The reaction solution was kept at −78° C. for 1 h then gradually warmed to RT. After completion of the reaction (checked by TLC), ice-cold 0.5 M HCl solution (70 ml) was added. The organic layer was separated and the aqueous layer was extracted with EtOAc (70 ml). The combined organic layer were washed with NaHCO₃ solution and brine, dried (Na₂SO₄), filtered and concentrated to give a crude product which was purified by silica gel chromatograph to yield compound 19 as a solid (3.5 g, 6.88 mmol, yield 55%). Alternative method for the preparation of compound 19:

Step 4b:

A mixture of BOC-L-alanine (17b) (2.9 g, 15.4 mmol), p-nitrophenol (3.3 g, 15.4 mmol) and DCC (3.3 g, 16.2 mmol) in EtOAc (60 ml) was stirred at RT for 2 h. A white precipitate was formed; the solid was filtered off and the filtrate was evaporated. The crude material was purified on a Biotage silica column, eluting with 20% hexane in CH₂Cl₂ to give compound 18b (2.8 g, 9 mmol, 58.4% yield) as a yellow solid. LCMS C₁₄H₁₈N₂O₆ [M+1]⁺311.1.

Step 5b: By a method analogous to that described in Example 5, Step 5, using compound 18b in place of compound 18, compound 19 was prepared.

Step 6: At 0° C., LiOH solution (150 mg, 6 mmol, in 15 ml of H₂O) was added to a solution of compound 19 (1.02 g, 2 mmol) in THF (37 ml). After 1 h at 0° C., the reaction was gradually warmed to RT and stirred at RT overnight. After the reaction was complete, EtOAc (50 ml) and H₂O (5 ml) were added, followed by the addition of 1 N HCl to acidify the mixture. The organic phase was separated, dried (Na₂SO₄), filtered and concentrated to give the title compound 20 as a white solid (0.94 g, 1.95 mmol, 98% yield). MS: C₂₂H₂₂F₃N₃O₆ [M+1]⁺482.1.

EXAMPLE 6

Step 1: A mixture of 4-chlorobenzaldehyde (21) (0.79 g, 5.45 mmol), 2-hydroxyethyl amine (22) (0.34 ml, 5.45 mmol) and Na₂SO₄ (1.44 g, 10.9 mmol) in dichloroethane (40 ml) was stirred at RT for 40 min. To this mixture, NaBH(OAc)₃ (3.12 g, 14.72 mmol) and AcOH (0.82 ml, 13.67 mmol) were added. After stirring at RT overnight, the reaction was quenched with saturated NaHCO₃ solution. The mixture was diluted with brine (200 ml) and extracted with CH₂Cl₂ (100 ml, 3×), combined and washed with brine (100 ml, 2×), dried (MgSO₄), filtered and evaporated to give crude compound 23 as an oil. The oil was purified with flash grade silica gel (100 g), eluting with 5% (1:9) (NH₄OH/CH₃OH)/95% CH₂Cl₂ to yield compound 23 (0.3 g, 1.62 mmol, 30% yield).

Step 2: A mixture of compound 20 (0.241 g, 0.5 mmol), compound 23 (92.8 mg, 0.5 mmol), HATU (285 mg, 0.75 mmol) and DIPEA (0.131 ml, 0.75 mmol) in dry DMF (3.0 ml) was stirred at RT for 4 h. After the reaction was complete, water (3 ml) was added to quench the reaction and the mixture was stirred for 10 min. Solid was collected, rinsed with water, and redissolved in CH₂Cl₂ (10 ml), dried (Na₂SO₄), filtered and evaporated. Product was purified by flash grade silica gel (100 g), eluting with 4.5% (1:9) (NH₄OH/CH₃OH)/95% CH₂Cl₂ to give pure compound 24 (0.18 g, 0.28 mmol, 56% yield) as a solid.

4 N HCl-dioxane solution (0.8 ml, 3.2 mmol) and CH₃OH (1 ml) were added to a solution of compound 24 (0.18 g, 0.33 mmol) in CH₂Cl₂ (2 ml). The mixture was stirred at RT overnight. Solvents were evaporated and product was triturated with CH₂Cl₂, filtered and dried under high vacuum to give title compound 25 as a HCl salt. LCMS: C₂₆H₂₄F₃N₄O₄Cl. HCl [M+1]⁺549.1

EXAMPLE 7

By employing methods analogous to those described in Example 6, Step 2, the following compounds were prepared using an appropriate aromatic or heteroaromatic amine coupled with compound 20 either by HATU or DEC (Example 4). The data for compounds of formula 26 are as follows:

Cpd. MS No. Structure (M + 1) 26-1

505 26-2

539 26-3

515 26-4

501 26-5

501 26-6

529 26-7

529 26-8

505 26-9

539 26-10

507 26-11

507 26-12

507 26-13

523 26-14

519 26-15

540 26-16

523 26-17

583 26-18

519 26-19

519 26-20

519 26-21

537 26-22

541 26-23

555 26-24

489 26-25

489 26-26

569 26-27

569 26-28

537 26-29

537 26-30

527 26-31

541 26-32

555 26-33

527 26-34

513 26-35

529 26-36

541 26-37

523 26-38

522 26-39

511 26-40

511 26-41

511 26-42

513 26-43

487 26-44

513 26-45

513 26-46

500 26-47

517 26-48

529 26-49

517 26-50

512 26-51

512 26-52

536 26-53

513 26-54

513 26-55

513 26-56

488 26-57

529 26-58

513 26-59

529 26-60

513 26-61

473 26-62

527 26-63

527 26-64

545 26-65

585 26-66

541 26-67

529 26-68

559 26-69

478 26-70

537 26-71

478 26-72

487 26-73

491 26-74

486 26-75

529 26-76

521 26-77

521 26-78

510 26-79

518 26-80

492 26-81

529 26-82

487 26-83

501 26-84

541 26-85

518 26-86

458 26-87

488 26-88

472 26-89

472 26-90

472 26-91

522 26-92

511 26-93

515 26-94

514 26-95

525 26-96

537 26-97

620 26-98

465 26-99

461 26-100

511 26-101

524 26-102

572 26-103

572 26-104

583 26-105

557 26-106

583 26-107

557 26-108

546 26-109

546 26-110

494 26-111

494 26-112

575 26-113

559 26-114

581 26-115

541 26-116

541 26-117

595 26-118

26-119

575 26-120

451 26-121

509 26-122

492 26-123

492 26-124

451 26-125

550 26-126

594 26-127

528 26-128

523 26-129

425 26-130

381 26-131

529 26-132

529 26-133

555 26-134

637 26-135

673 26-136

588 26-137

556 26-138

566 26-139

588 26-140

556 26-141

494 26-142

425 26-143

541 26-144

381 26-145

528 26-146

563 26-147

479 26-148

479 26-149

536 26-150

549 26-151

572 26-152

583 26-153

601 26-154

558 26-155

554 26-156

639 26-157

589 26-158

604 26-159

618 26-160

541 26-161

541 26-162

541 26-163

542 26-164

604 26-165

604 26-166

541 26-167

555 26-168

584 26-169

521 26-170

535 26-171

594 26-172

543 26-173

608 26-174

536 26-175

586 26-176

543 26-177

542 26-178

507 26-179

507 26-180

507 26-181

509 26-182

543 26-183

556 26-184

562 26-185

514 26-186

589 26-187

519 26-188

556 26-189

493 26-190

562 26-191

515 26-192

558 26-193

546 26-194

526 26-195

618 26-196

560 26-197

572 26-198

508 26-199

533 26-200

523 26-201

571 26-202

572 26-203

556 26-204

583 26-205

538 26-206

522 26-207

573 26-208

550 26-209

555 26-210

550 26-211

556 26-212

536 26-213

525 26-214

525 26-215

509 26-216

608 26-217

583 26-218

583 26-219

560 26-220

540 26-221

473 26-222

571 26-223

507 26-224

605 26-226

620 26-227

577 26-228

557 26-229

568 26-230

569 26-231

556 26-232

542 26-234

558 26-235

558 26-236

548 26-237

572 26-238

543 26-239

538 26-240

614 26-241

615 26-242

520 26-243

630 26-244

571 26-245

543 26-246

534 26-247

552 26-248

527 26-249

537 26-250

582 26-251

556 26-252

547 26-253

533 26-254

586 26-255

516 26-256

502 26-257

545 26-258

564 26-259

557 26-260

540 26-261

544 26-262

595 26-263

542 26-264

555 26-265

546 26-266

557 26-267

546 26-268

542 26-269

556 26-270

537 26-271

551 26-272

571 26-273

527 26-274

439 26-275

552 26-276

548 26-277

552 26-278

586 26-279

583 26-280

561 26-281

532 26-282

439 26-283

532 26-264

533 26-285

608 26-286

620 26-287

560 26-288

546 26-289

512 26-290

494 26-291

508 26-292

474 26-293

518 26-294

439 26-295

585 26-296

546 26-297

546 26-298

569 26-299

594 26-300

564 26-301

561 26-302

583 26-303

572 26-304

530 26-305

529 26-306

506 26-307

530 26-308

556 26-309

542 26-310

453 26-311

506 26-312

556 26-313

508 26-314

554 26-315

532 26-316

506 26-317

507 26-318

586 26-319

552 26-320

560 26-321

506 26-322

492 26-323

516 26-324

642 26-325

532 26-326

466 26-327

537 26-328

600 26-329

531 26-330

480 26-331

466 26-332

453 26-333

466 26-334

522 26-335

516 26-336

551 26-337

514 26-338

632 26-339

519 26-340

516 26-341

508 26-342

508 26-343

516 26-344

496(M +! +H) 26-345

439 26-346

574 26-347

590 26-348

536 26-349

542 26-350

627 26-351

582 26-352

534 26-353

542 26-354

542 26-355

542 26-356

532 26-357

532 26-358

600 26-359

600 26-360

550 26-361

556 26-362

542 26-363

528 26-364

528 26-365

528 26-366

570 26-367

542 26-368

532 26-369

532 26-370

533 26-371

533 26-372

614 26-373

560 26-374

492 26-375

548 26-376

548 26-377

534 26-378

532 26-379

654 26-380

521 26-381

590 26-382

522 26-383

562 26-384

530 26-385

586 26-386

514 26-387

521 26-388

605 26-389

592 26-390

606 26-391

494 26-392

494 26-393

542 26-394

544 26-395

530 26-396

575 26-397

629 26-398

520 26-399

615 26-400

542 26-401

592 26-402

516 26-403

600 26-404

564 26-405

542 26-406

571 26-407

536 26-408

536 26-409

506 26-410

492 26-411

492 26-412

478 26-413

554 26-414

600 26-415

542 26-416

465 26-417

465 26-418

536 26-419

536 26-420

509 26-421

520 26-422

509 26-423

520 26-424

506 26-425

451 26-426

504 26-427

530 26-428

506 26-429

504 26-430

520 26-431

547 26-432

528 26-433

518 26-434

532 26-435

592 26-436

536 26-437

551 26-438

550 26-439

518 26-440

528 26-441

614 26-442

521 26-443

520 2-444

506 26-445

619 26-446

580 26-447

542 26-448

557 26-449

590 26-450

506 26-451

518 26-452

522 26-453

481 26-454

481 26-455

467 26-456

467 26-457

473 26-458

481 26-459

530 26-460

522 26-461

588 26-462

534 26-463

520 26-464

541 26-465

535 26-466

541 26-467

549 26-468

501 26-469

548 26-470

507 26-471

509 26-472

511 26-473

531 26-474

557 26-475

534 26-476

520 26-477

590 26-478

525 26-479

537 26-480

589 26-481

540 26-482

473 26-483

502 26-484

587 26-485

520 26-486

543 26-487

551 26-488

527 26-489

423 26-490

437 26-491

421 26-492

435 26-493

449 26-494

435 26-495

479 26-496

435 26-497

463 26-498

465 26-499

466 26-500

487 26-501

511 26-502

 541,543 26-503

531 26-504

492 26-505

483 26-506

512 26-507

557 26-508

455 26-509

549 26-510

518 26-511

520 26-512

475 26-513

461 26-514

541 26-515

524 26-516

501 26-517

549 26-518

461 26-519

503 26-520

517 26-521

549 26-522

546 26-523

527 26-524

521 26-525

528 26-526

528 26-527

463 26-528

451 26-529

451 26-530

437 26-531

465 26-532

451 26-533

497 26-534

552 26-535

532 26-536

546 26-537

556 26-538

526 26-539

536 26-540

534 26-541

518 26-542

520 26-543

479 26-544

479 26-545

451 26-546

451 26-547

437 26-548

521 26-549

528 26-550

529 26-551

497 26-552

511 26-553

502 26-554

479 26-555

445 26-556

449 26-557

423 26-558

437 26-559

479 26-560

409 26-561

479 26-562

562 26-563

501 26-564

479 26-565

493 26-566

395 26-567

409 26-568

537 26-569

555 26-570

529 26-571

487 26-572

559 26-573

549 26-574

549 26-575

478 26-576

572 26-577

499 26-578

499 26-579

477 26-580

423 26-581

546 26-582

534 26-583

492 26-584

492 26-585

548 26-586

569 26-587

583 26-588

501 26-589

437 26-590

451 26-591

437 26-592

437 26-593

481 26-594

451 26-595

481 26-596

520 26-597

485 26-598

546 26-599

463 26-600

492 26-601

506 26-602

478 26-603

492 26-604

506 26-605

492 26-606

506 26-607

518 26-608

569 26-609

547 26-610

545 26-611

545 26-612

529 26-613

457 26-614

501 26-615

443 26-616

492 26-617

545 26-618

532 26-619

532 26-620

504 26-621

492 26-622

531 26-623

585 26-624

534 26-625

534 26-626

479 26-627

479 26-628

535 26-629

508 26-630

457 26-631

439 26-632

453 26-633

492 26-634

520 26-635

465 26-636

489 26-637

613 26-638

522 26-639

463 26-640

449 26-641

463 26-642

538 26-643

538 26-644

447 26-645

463 26-646

627 26-647

588 26-648

554 26-649

554 26-650

451 26-651

465 26-652

527 26-653

560 26-654

560 26-655

476 26-656

526 26-657

518 26-658

528 26-659

517 26-660

506 26-661

506 26-662

508 26-663

524 26-664

541 26-665

548 26-666

520 26-667

554 26-668

529 26-669

464 26-670

465 26-671

478 26-672

492 26-673

395 26-674

508 26-675

546 26-676

532 26-677

571 26-678

514 26-679

588 26-680

519 26-681

570 26-682

514 26-683

518 26-684

528 26-685

540 26-686

511 26-687

520 26-688

548 26-689

546 26-690

528 26-691

528 26-692

521 26-693

520 26-694

506 26-695

453 26-696

565 26-697

504 26-698

520 26-699

534 26-700

503 26-701

544 26-702

421 26-703

547 26-704

492 26-705

467 26-706

451 26-707

570 26-708

542 26-709

484 26-710

569 26-711

574 26-712

491 26-713

572 26-714

558 26-715

548 26-716

564 26-717

565 26-718

551 26-719

497 26-720

542 26-721

492 26-722

506 26-723

532 26-724

546 26-725

518 26-726

534 26-727

565 26-728

564 26-729

564 26-730

506 26-731

532M + Na 26-732

535 26-733

557 26-734

520 26-735

568 26-736

554 26-737

467 26-738

540 26-739

506 26-740

518 26-741

558 26-742

498 26-743

498 26-744

493 26-745

465 26-746

527 26-747

507 26-748

556 26-749

556 26-750

572 26-751

556 26-752

695 26-753

556 26-754

594 26-755

533 26-756

547 26-757

549 26-758

527 26-759

554 26-760

636 26-761

619 26-762

699 26-763

549 26-764

527 26-765

506 26-766

518 26-767

526 26-768

526 26-769

592 26-770

654 26-771

496 26-772

598 26-773

510 26-774

529 26-775

487 26-776

485 26-777

584 26-778

584 26-779

508 26-780

563 26-781

596 26-782

473 26-783

557 26-784

570 26-785

564 26-786

534 26-787

548 26-788

540 26-789

530 26-790

529 26-791

522 26-792

536 26-793

550 26-794

562 26-795

479 26-796

532 26-797

548 26-798

562 26-799

530 26-800

478 26-801

570 26-802

554 26-803

522 26-804

536 26-805

548 26-806

576 26-807

554 26-808

546 26-809

550 26-810

520 26-811

598 26-812

598 26-813

572 26-814

492 26-815

492 26-816

497 26-817

506 26-818

489 26-819

539 26-820

570 26-821

570 26-822

575 26-823

531 26-824

580 26-825

558 26-826

506 26-827

515 26-828

557 26-829

465 26-830

566 26-831

507 26-832

514 26-833

531 26-834

496 26-835

510 26-836

580 26-837

518 26-838

532 26-839

527 26-840

514 26-841

510 26-842

497 26-843

555 26-844

517 26-845

532 26-846

606 26-847

584 26-848

584 26-849

518 26-850

532 26-851

462 26-852

476 26-853

463 26-854

536 26-855

534 26-856

512 26-857

519 26-858

524 26-859

538 26-860

522 26-861

584 26-862

584 26-863

569 26-864

569 26-865

546 26-866

551 26-867

573 26-868

558 26-869

559 26-870

586 26-871

552 26-872

479 26-873

546 26-874

478 26-875

506 26-876

527 26-877

558 26-878

514 26-879

544 26-880

544 26-881

572 26-882

534 26-883

516 26-884

584 26-885

518 26-886

534 26-887

465 26-888

529 26-889

496 26-890

572 26-891

522 26-892

478 26-893

493 26-894

580 26-895

493 26-896

527 26-897

558 26-898

558 26-899

490 26-890

487 26-891

518 26-892

544 26-893

594 26-894

612 26-895

560 26-896

528 26-897

540 26-898

487 26-899

515 26-900

501 26-901

511 26-902

529 26-903

545 26-904

551 26-905

617 26-906

599 26-907

560 26-908

574 26-909

501 26-910

529 26-911

564 26-912

578 26-913

499

EXAMPLE 8

-   Step 1:

Using N-[(1,1-dimethylethoxy)carbonyl]-O-(1,1-dimethylethyl)-L-serine (compound 27) as starting material, [1(S)-[(1,1-dimethylethoxy)methyl]-2-fluoro-2-oxoethyl] carbamic acid, 1,1-dimethylethyl ester (compound 28) was prepared by a method analogous to that in Example 5, step 4.

-   Step 2:

0.5 M KHMDS in toluene (92.5 ml, 46.25 mmol) was added slowly via a syringe to a mixture of compound 16 (8.5 g, 22 mmol) and compound 28 (6.8 g, 25.8 mmol) in dry THF (90 ml) at −78° C. The mixture was slowly warmed to RT, then stirred at RT for 1 h. After the reaction was complete, it was quenched with 1 N HCl (80 ml)(cooled with ice-water bath), diluted with saturated NH₄Cl solution (100 ml), extracted with EtOAc (200 ml×2), dried (Na₂SO₄), filtered and evaporated. Crude material was purified on Biotage with CH₂Cl₂ (4 L) and 5% EtOAc/CH₂Cl₂ (4 L) to give compound 29 as a light yellow solid (6.5 g, 11.8 mmol, 52%). MS C₂₈H₃₄F₃N₃O₇ [M+1]⁺582.1.

EXAMPLE 9

Compound 29 (13.5 g, 23.24 mmol) was treated with THF:H₂O (2:1) (200 ml) and LiOH.H₂O (0.95 g, 39.6 mmol) (dissolved in 10 ml of H₂O). After stirring at RT for 2 h, the suspension was not dissolved. Additional THF:H₂O (2:1) (100 ml) and LiOH.H₂O (0.95 g, 39.6 mmol) was added. It was stirred at RT overnight. After completion, the reaction was neutralized with 1 N HCl. The mixture was extracted with CH₂Cl₂ (100 ml×3), combined, washed with brine (100 ml), dried (Na₂SO₄), filtered and evaporated to give the title compound 30 as a yellow solid (11.8 g, 21.3 mmol, 92%). LCMS: C₂₆H₃F₃N₃O₇ [M+1]⁺554.1.

EXAMPLE 10

-   Step 1:

By a method analogous to Example 2, using 5-fluoro-3-methyl-benzo[B]-thiophene (31) as starting material, compound 33 was obtained. It was treated with 10 equivalents of 98% hydrazine in absolute EtOH and CH₂Cl₂ (1:1) to give compound 34, which was purified by treatment with a slight excess of 4 N HCl/dioxane solution to give compound 35 as a HCl salt. FABMS: C₉H₈FNS. HCl [M+1]⁺182.0

-   Step 2:

By methods analogous to those described in Example 3, using compound 35 as a starting material, compound 36 was obtained.

-   Step 3:

The protecting groups on compound 36 were removed by treatment with HCl-dioxane/CH₂Cl₂ or CF₃COOH. The title compound 37 was obtained directly as a HCl salt or as a TFA salt depending on the acid treatment. The TFA salt was neutralized with NH₄OH and converted to HCl salt with 1.0 equivalent of HCl. HRMS C₂₆H₂₀F₄N₄O₄S. HCl calculated [M+1]⁺561.1220, Found 561.1230.

EXAMPLE 11

By employing analogous methods to those described in Example 10, the following compounds were obtained as HCl salts using compound 30 coupled with the appropriate primary or secondary amine, followed by removal of the protecting group as described for Example 10, step 3.

Cpd. MS No. Structure (M + 1) 38-1

521 38-2

557 38-3

523 38-4

539 38-5

523 38-6

523 38-7

555 38-8

539 38-9

513 38-10

529 38-11

535 38-12

505 38-13

505 38-14

537 38-15

529 38-16

543 38-17

601 38-18

561 38-19

529 38-20

541 38-21

545 38-22

553 38-23

532 38-24

558 38-25

527 38-26

604 38-27

557 38-28

636 38-29

557 38-30

397 38-31

590 38-32

557 38-33

573 38-34

553 38-35

548 38-36

562

EXAMPLE 12

By employing methods analogous to those described for Example 5, using compound 18c in place of compound 18, compound 42 was obtained, which was treated with LiOH.H₂O to give the title compound 43.

EXAMPLE 13

By employing methods analogous to those described in Example 6, using compound 43 in place of compound 20 and 4-chlorobenzylamine in place of compound 23 in the coupling reaction, compound 44 was obtained. After removal of the t-BOC group of compound 44 with HCl, the title compound 45 was obtained as a HCl salt. MS: C₂₅H₂₂ClF₃N₄O₃. HCl [M+1]⁺519.1.

Using a procedure similar to that described for compound 45, the following compounds were prepared:

Cpd. MS No. Structure (M + 1) 45-1

562 45-2

479 45-3

557 45-4

541 45-5

576 45-6

569 45-7

506 45-8

479 45-9

449 45-10

435 45-11

506

EXAMPLE 14

By employing methods analogous to those described in Example 6, using compound 43 in place of compound 20 and compound 9 in place of compound 23 compound 46 was obtained. After removal of the t-BOC group of compound 46 with HCl, the title compound 47 was obtained as a HCl salt. MS C₂₇H₂₃F₃N₄O₃S .HCl [M+1]⁺541.1.

EXAMPLE 15

By employing methods analogous to those described in Example 5, using compound 18d in place of compound 18, compound 48 was obtained, which was treated with LiOH.H₂O to obtain the title compound 49.

EXAMPLE 16

By employing methods analogous to those described in Example 6, using compound 50 in place of compound 20 and 2,4-diflurobenzylamine in place compound 23, compound 51 was obtained. After removal of the t-BOC group of compound 51 with HCl, the title compound 52 was obtained as a HCl salt. MS: C₂₆H₂₃F₅N₄O₃. HCl [M+1]⁺535.

Using similar procedures and the appropriate staring materials, the following compounds were also prepared:

Cpd. MS No. Structure (M + 1) 47-1

542 47-2

541 47-3

519 52-1

556

EXAMPLE 17

By employing methods analogous to those described in Example 5, using compound 18e in place of compound 18, compound 53 was obtained, which was treated with LiOH.H₂O to yield the title compound 54.

EXAMPLE 18

By employing methods analogous to those described in Example 10, using compound 55 in place of compound 30 and 1-amino-2-hydroxyethyl benzene in place of compound 35, compound 56 was obtained. After purification and removal of the t-BOC group of compound 56 with HCl, compound 57 was obtained as a HCl salt. MS: C₂₆H₂₅F₃N₄O₅. HCl [M+1]⁺567.1.

EXAMPLE 19

Using the appropriate aromatic or heteroaromatic amine reagent coupled with compound 55 according to the procedure described for Example 10, steps 2 and 3, the desired compound 58 was obtained as a hydrochloride salt.

Cpd. MS No. Structure (M + 1) 58-1

537 58-2

564 58-3

543

EXAMPLE 20

-   Step 1: DBU (1.7 g, 11 mmol) was added to a mixture of     4-chromanol (59) (1.5 g, 10 mmol) and diphenyl-phosphoryl azide     (DPPA) (3.0 g, 11 mmol) in CH₂Cl₂ (10 ml) at RT. The mixture     immediately turned brown (a water bath was used to cool the reaction     temperature). The solution was stirred at RT overnight. After     completion of the reaction, the reaction mixture was diluted with     ether/EtOAc (1:1) (100 ml) and washed with saturated NaHCO₃, 5% HCl     and brine. The organic layer was dried (Na₂CO₃), filtered and     concentrated to give a residue which was purified by column     chromatography, eluting with 30% CH₂Cl₂/hexane to give compound 60     (1.45 g, 0.83 mmol) with a 83% of yield. -   Step 2: 4 N HCl/dioxane (2 ml) and 10% Pd/C (0.5 g) were added to a     solution of compound 60 (1.3 g, 7.4 mmol) in MeOH (50 ml). The     mixture was stirred under a H₂ balloon at RT for 46 h. After the     reaction was complete, the solid was filtered off. The filtrate was     concentrated and to obtain the desired amine (61) as a light yellow     HCl salt. LCMS C₉H₁₁NO. HCl [M+1]⁺149.0.

EXAMPLE 21

By employing methods analogous to those described for Example 20, replacing compound 59 with 62 and 60 with 63, the title compound 64 was obtained as an amine HCl salt. LCMS C₉H₁₁O₂N. HCl [M+1]⁺166.0

EXAMPLE 22

NaBH₄ (0.7 g, 18.5 mmol) at RT was added to a solution of compound 65 (0.6 g, 3.57 mmol) in MeOH (20 ml) cooled with a water bath. After 10 min, solvent was removed. The residue was treated with 5% NaHCO₃ and the product was extracted with CH₂Cl₂, then with EtOAc. The combined organic solution was washed with brine, dried (Na₂SO₄), filtered and concentrated to give compound 66 as a white solid. Using a method similar to Example 27, the title compound 67 was obtained as a HCl salt. LCMS C₇H₁₁N₃O₂.HCl [M+1]⁺170.0

EXAMPLE 23

Solid 1,3,5-triazine (2.4 g, 30 mmol) (68) was mixed with 1-(pyrrolidino)-1-cyclohexene (69) in a pressure tube (15 ml) and heated at 93° C. (bath temperature) with stirring for 22 h. After completion of the reaction, the reaction mixture was concentrated and dissolved in CH₂Cl₂, washed with saturated NaHCO₃ solution, dried (Na₂SO₄), then purified by column chromatography to give compound 70 as a solid.

The title compound 73 was prepared according to the procedure described for Example 2, and step 2 of Example 3.

EXAMPLE 24

A mixture containing 3-acetylthianaphthene (74) (0.5 g, 2.8 mmol), allylamine (0.42 g, 5.6 mmol), NaBH(OAc)₃ (1.2 g, 5.6 mmol), and HOAc (0.15 ml) in dichloroethane (15 ml) was stirred at RT overnight. After completion, the reaction mixture was quenched with NaHCO₃ and extracted with CH₂Cl₂. The organic solvent was dried (Na₂SO₄), filtered and evaporated. The crude material was purified by column chromatography to give compound 75 (0.43 g) as an oil.

N,N-dimethylbarbituric acid (0.65 g, 4.14 mmol) and tetrakis-(triphenyl-phosphine)palladium (16 mg, 0.0138 mmol) were added to a solution of compound 75 (0.3 g, 1.38 mmol) in CH₂Cl₂ (30 ml). The mixture was stirred at 40° C. for 2 h, then at RT overnight. After completion of the reaction, the mixture was diluted with CH₂Cl₂ and washed with saturated Na₂CO₃ solution. The organic layer was separated and the aqueous layer was re-extracted with CH₂Cl₂. The organic fractions were combined and concentrated. The crude material was purified by silica gel chromatography to give the title compound 76 as an oil.

EXAMPLE 25

-   Step 1: MgO (0.4 g, 10 mmol) and 10% Pd/C (0.5 g) were added to a     solution of compound 77 (1.0 g, 5.4 mmol) in EtOH:MeOH (1:1) (100     ml). The mixture was stirred at RT overnight. After completion of     the reaction, MgO and Pd/C were filtered off and the filtrate was     concentrated to dryness. The residue was dissolved in EtOAc and     washed with water, dried (Na₂SO₄), filtered and evaporated to give a     solid compound 78. -   Step 2: Using the procedure described in Example 2, Step 1, the     bromo derivative 79 was obtained. The crude material was used in the     next reaction without purification. -   Step 3: Sodium azide was mixed with DMSO and stirred at RT until all     solid was dissolved. Compound 78 was added at RT, and after stirring     at RT for 1 h, ice-water was added. The product was extracted with     EtOAc:ether (1:1). The combined organic layers were washed with     water, dried (Na₂SO₄), filtered and evaporated to obtain the azido     derivative 80, as an oil. -   Step 4: By employing methods analogous to Example 20, Step 2, the     azido derivative 80 was converted to the title compound 81 as a     hydrochloride salt.

EXAMPLE 26

-   Step 1: Znl₂ (0.64 g, 2 mmol) was added in one portion at RT under     N₂ to a mixture of 2,4-dichlorobenzaldehyde (82) (3.5 g, 20 mmol)     and TMSCN (2.6 g, 26 mmol). After 15 min, 7 N NH₃ solution in MeOH     (20 ml) was added and the mixture was stirred at 40° C. for 2 h.     Solvents were evaporated and the residue was re-dissolved in Et₂O,     washed with water, dried (MgSO₄) and filtered. HCl gas was bubbled     through the filtrate to give an off-white solid of compound 83 (3.5     g, 74%). MS: C₈H₆N₂Cl₂, [M+1]⁺202. -   Step 2: A stream of HCl gas was bubbled through a solution of     compound 83 (3.5 g, 14.8 mmol) in MeOH (85 ml) for 4 h. Water (2 ml)     was added and the reaction mixture was concentrated to provide an     off-white solid of compound 84 (3.4 g, 90%) as a HCl salt. MS:     C₈H₈N₂OCl₂, [M+1]⁺219

EXAMPLE 27

-   Step 1: Compound 85 (2.4 g, 14.4 mmol) was suspended in phosphorus     oxychloride (30 ml) and heated to reflux for 15 h. The reaction     mixture was cooled to RT, and saturated (aq) NaHCO₃ (250 ml) was     added carefully with vigorous stirring at 0° C., followed by the     addition of Et₂O (150 ml). The aqueous layer was separated and     extracted with Et₂O. The organic layers were combined, washed with     brine, filtered and concentrated to give compound 86 as a yellow oil     (2.14 g, 81%). MS: C₇H₅ClN₂S [M+1] 185; [M+2], 186 -   Step 2: Compound 87 was obtained as a white solid (93% yield)     according to the procedure described for the Example 25, Step 1. MS:     C₇H₆N₂S [M+1]⁺151. -   Step 3: Compound 88 [MS: C₇H₅BrN₂S [M+1]+Br⁷⁹ 229, Br⁸¹ 231] was     synthesized from compound 88 according to the method described in     Example 2, Step 1. The bromo-derivative 88 was converted to its     azido-derivative-89 [MS: C₇H₅N₅S [M+1]+192] according to the     procedure described for Example 25, Step 3. The title compound 90     was obtained as a HCl salt by hydrogenation of compound 89 according     to the procedure described for Example 20. MS: C₇H₇N₃S [M+1]⁺166.

EXAMPLE 28

-   Step 1: Compound 91 (1.47 g, 7.99 mmol) was treated with a solution     of 0.5 M NaOMe in MeOH (32 ml) under N₂. The suspension was stirred     at RT overnight. The solvent was removed and the resultant residue     partitioned between EtOAc (75 ml) and water (75 ml). The aqueous     layer was separated and extracted with EtOAc. The organic extracts     were combined, dried (MgSO₄), filtered and concentrated in vacuo to     give 1.41 g (98%) of white solid, compound 92. MS: C₈H₈N₂OS     [M+1]⁺181. -   Step 2: The title compound 93 was obtained from compound 92 using     methods similar to those described for Example 27, Step 3. MS:     C₈H₉N₃SO [M+1]⁺196.

EXAMPLE 29

Compound 94 was prepared according to the literature (Tetrahedron Letters 41, 8661-8664, (2000)). Compound 94 (3.0 g, 22.5 mmol) was mixed with NaI (3.37 g, 22.5 mmol) and NaN₃ (1.9 g, 29 mmol) in CH₂Cl₂/acetone (1:1, 250 ml) and refluxed for 36 h. After completion of the reaction, the reaction mixture was filtered and the filtrate concentrated to dryness. Crude compound 95 was purified on Biotage system, eluting with 2% MeOH in CH₂Cl₂, to obtain pure compound 95 as a white solid. Compound 95 (2.46 g, 17.6 mmol) was dissolved in MeOH (100 ml) and formic acid (0.81 ml, 17.6 mmol), and 10% Pd/C (490 mg) was added. The mixture was stirred at RT under a H₂ balloon overnight. The solids were filtered off and the filtrate was evaporated to give the title compound, 96, as a formic acid salt. MS: C₃H₆N₄O [M+1]⁺115.

EXAMPLE 30

-   Step 1: To a solution of compound 97 (280 mg, 0.462 mmol) dissolved     in THF (10 ml) was added Lawesson reagent (467 mg, 1.15 mmol). The     reaction mixture was heated at reflux for 24 h then cooled to RT.     The solvent was evaporated. Purification by silica gel     chromatography (eluant: 1%-3% EtOAc—CH₂Cl₂) gave 166 mg (0.267 mmol,     58%) of the product 98 as a yellow foam. MS (M+1): m/e 623. -   Step 2: To a solution of compound 98 (273 mg, 0.438 mmol) dissolved     in CH₂Cl₂ (4 ml) was added TFA (1 ml). The reaction mixture was     stirred at RT for 4 h. The solvent was evaporated, and the crude     product was dissolved in 10 ml of 1:1 CH₂Cl₂: MeOH and     diethylaminomethylpolystyrene resin (0.50 g, from Fluka) was added.     The resulting mixture was stirred for 15 min, filtered, and the     resin was washed with MeOH. The filtrate was evaporated.     Purification by silica gel chromatography (eluant: 2%-3%     MeOH—CH₂Cl₂) gave 169 mg (0.323 mmol, 74%) of the product 99 as a     yellow foam. MS (M+1): m/e 523.

EXAMPLE 31

-   Step 3: To a solution of trans-4-benzoyl-cyclohexylamine (2.06 g,     9.4 mmol) dissolved in dry CH₂Cl₂ (50 ml) was added 3A sieves (3 g),     Et₃N (2.38 g, 3.3 ml, 23.5 mmol),     [4-(N-BOC-aminomethyl)phenyl]boronic acid 100A (3.00 g, 11.9 mmol),     and copper acetate (2.16 g, 11.9 mmol). The reaction mixture was     stirred at RT for 24 h. 2 N aqueous NH₄OH (50 ml) was added, and the     reaction mixture was filtered to remove the sieves which were washed     with additional CH₂Cl₂ and 2 N aqueous NH₄OH. The layers of the     filtrate were separated, and the aqueous layer was extracted width     CH₂Cl₂. The combined organic extract was dried (MgSO₄), filtered,     and concentrated. Purification by silica gel chromatography (eluant:     5%-10% EtOAc—CH₂Cl₂) gave 0.73 g (1.72 mmol, 18%) of the product     101A as a yellow foam. MS (M+1): m/e 425.

The following intermediates were synthesized by using a similar procedure:

Number Compound MS (M + 1) 101B

277 101C

371 101D

349 101E

321 101F

335 101G

335 101H

425 101I

277 101J

371 101K

411 101L

335 101M

335

-   Step 2: To a solution of compound 101A (0.72 g, 1.70 mmol) dissolved     in THF (6 ml) MeOH (6 ml), and water (3 ml) was added LiOH (0.36 g,     8.48 mmol). The reaction mixture was stirred at RT for 4 h. The     solvent was evaporated, saturated NH₄Cl (25 ml) was added, and the     aqueous solution was extracted with CH₂Cl₂. The combined organic     extract was dried (MgSO₄), filtered, and concentrated. Purification     by silica gel chromatography (eluant: 3%-5% MeOH—CH₂Cl₂) gave 0.48 g     (1.50 mmol, 89%) of the product 102A as a white foam. MS (M+1): m/e     321.

The following intermediates were synthesized by using a similar procedure:

MS Number Compound (M + 1) 102B

267 102C

307 102D

279 102E

293 102F

293 102G

321 102H

267 102I

307 102J

293 102K

293

-   Step 3: To a solution of compound 102A (475 mg, 1.48 mmol) dissolved     in 1:1 CH₂Cl₂:MeOH (10 ml) was added 4 N HCl in dioxane (3.0 ml,     11.9 mmol). The reaction mixture was stirred at RT for 3 h. The     solvent was evaporated to give 429 mg (1.46 mmol, 99%) of the     product 103A as a white solid. MS (M+1): m/e 221.

To a solution of compound 102D (0.73 g, 2.62 mmol) suspended in CH₂Cl₂ (18 ml) was added TFA (3 ml). The reaction mixture was stirred at RT for 3 h. The solvent was evaporated, and the TFA salt of the product was dissolved in MeOH (20 ml). Diethylaminomethylpolystrene resin (4 g, Fluka) was added and stirred at RT for 20 min. The resin was removed by filtration and washed with MeOH. The filtrate was concentrated to give 0.47 g (2.62 mmol, 100%) of the product 103B as a yellow solid. MS (M+1-OH): m/e 162.

The following intermediates were synthesized by using a similar procedure:

Number Compound MS (M + 1) 103C

150(M + 1-OH) 103D

207 103E

176M + 1-OH 103F

176M + 1-OH 103G

221 103H

167 103I

207 103J

193 103K

193

EXAMPLE 32

-   Step 1: To a solution of methyl 4-(BOC-aminomethyl)-benzoate 104A     (4.15 g, 15.6 mmol) dissolved in THF (20 ml), MeOH (20 ml), and     water (10 ml) was added LiOH (0.72 g, 17.2 mmol). The reaction     mixture was stirred at RT for 24 h. The solvent was evaporated to     give 4.02 g (15.6 mmol, 100%) of the product 105A as a white solid.     MS (M+2-tBu for acid COOH): m/e 196. -   Step 2: To a solution of 4-hydroxypiperidine (0.41 g, 4.08 mmol)     dissolved in dry DMF (20 ml) was added 3A sieves (1.0 g) and the     mixture was stirred at RT for 15 min. HOBT (0.55 g, 4.08 mmol), EDCI     (0.78 g, 4.08 mmol), compound 105A (0.70 g, 2.72 mmol), and Et₃N     (0.55 g, 0.76 ml, 5.44 mmol) were then added. The reaction mixture     was stirred at RT for 20 h. The solvent was evaporated, 0.2 N NaOH     (40 ml) was added, and the aqueous solution was extracted with     CH₂Cl₂. The combined organic extract was dried (MgSO₄), filtered,     and concentrated. Purification by silica gel chromatography (eluant:     5%-10% MeOH—CH₂Cl₂) gave 0.78 g (2.33 mmol, 86%) of the product 106A     as a white foam. MS (M+1): m/e 335.

The following intermediates were synthesized by using a similar procedure:

MS Number Compound (M + 1) 106B

307 106C

321 106D

321 106E

335 106F

307 106G

321 106H

321 106I

266 106J

308 106K

306 106L

322

-   Step 3: Using the procedure of step 3 from Example 31, the following     intermediates were synthesized:

Number Compound MS (M + 1) 107A

235 107B

207 107C

221 107D

221 107E

235 107F

207 107G

221 107H

221 107I

166 107J

208 107K

206 107L

222

-   Step 4: To a solution of compound 104A (2.47 g, 9.31 mmol) dissolved     in Et₂O (50 ml) was added LiBH₄ (0.81 g, 37.2 mmol) then MeOH (1.19     g, 1.5 ml, 37.2 mmol). The reaction mixture was heated at reflux for     5 h and then cooled to RT. The solvent was evaporated. Water (50 ml)     was added, and the aqueous solution was extracted with CH₂Cl₂. The     combined organic extract was dried (MgSO₄), filtered, and     concentrated. Purification by silica gel chromatography (eluant:     5%-8% MeOH —CH₂Cl₂) gave 2.15 g (9.06 mmol, 97%) of the product 108A     as a white solid. MS (M+1): m/e 238.

The following intermediate was synthesized by using a similar procedure:

Number Compound MS (M + 1) 108B

182M + 2-tBu

-   Step 5: To a solution of oxalyl chloride (1.43 g, 0.98 ml, 11.3     mmol) dissolved in dry CH₂Cl₂ (20 ml) and cooled to −78° C. under a     N₂ atmosphere was added DMSO (1.76 g, 1.6 ml, 22.5 mmol) dissolved     in CH₂Cl₂ (5 ml) dropwise via addition funnel. The reaction mixture     was stirred at −78° C. for 15 min then compound 108A (2.14 g, 9.02     mmol) dissolved in CH₂Cl₂ (25 ml) was added dropwise via addition     funnel. The reaction mixture was stirred at −78° C. for 60 min, then     Et₃N (2.74 g, 3.8 ml, 27.0 mmol) was added. The reaction mixture was     stirred at −78° C. for 20 min, then warmed to RT. Water (75 ml) was     added, and the aqueous solution was extracted with CH₂Cl₂. The     combined organic extract was dried (MgSO₄), filtered, and     concentrated. Purification by silica gel chromatography (eluant:     2%-3% MeOH —CH₂Cl₂) gave 2.12 g (9.02 mmol, 100%) of the product     109A as a white solid. MS (M+1): m/e 236.

The following intermediate was synthesized by using a similar procedure:

Number Compound MS (M + 1) 109B

180M + 2-tBu

-   Step 6: To a solution of compound 109A (0.50 g, 2.12 mmol) dissolved     in 10% water by volume in EtOH (20 ml) was added sodium acetate     (1.05 g, 12.7 mmol) and hydroxylamine hydrochloride (0.59 g, 8.50     mmol). The reaction mixture was heated at reflux for 4 h and then     cooled to RT. The solvent was evaporated. Water (30 ml) was added,     and the aqueous solution was extracted with CH₂Cl₂. The combined     organic extract was dried (MgSO₄), filtered, and concentrated to     give 0.53 g (2.12 mmol, 100%) of the product 110A as a white solid.     MS (M+1): m/e 251.

The following intermediate was synthesized by using a similar procedure:

Number Compound MS (M + 1) 110B

195M + 2-tBu

-   Step 7: Using the procedure of step 3 from Example 31, the following     intermediates were synthesized:

Number Compound MS (M + 1) 111A

151 111B

151

EXAMPLE 33

-   Step 1: 3-Methylphthalic anhydride 112A (5.00 g, 30.8 mmol) and urea     (1.85 g, 30.8 mmol) were combined and heated at 320-350° C. with     stirring for 5 min, then cooled to RT. The brown solid was     triturated with water and filtered. The solid was washed with water     and dried to give 4.80 g (29.8 mmol, 97%) of the product 113A as a     pink solid. MS (M+1): m/e 162.

The following intermediates were synthesized by using a similar procedure:

Number Compound MS (M + 1) 113B

204 113C

164 113D

165 forM+ 113E

184

-   Step 2: To compound 113A (4.80 g, 29.8 mmol) was added 1 M borane in     THF (104 ml, 0.104 mol) under a N₂ atmosphere. The reaction mixture     was heated at reflux for 16 h and then cooled to 0° C. EtOH (80 ml)     and K₂CO₃ (9.20 g, 66 mmol) were added carefully. The resulting     mixture was heated at reflux for 16 h and then cooled to RT.     (tBOC)₂O (10.00 g, 45.8 mmol) was added, and the reaction mixture     was stirred at RT for 3 h. The solvent was evaporated. Water     (200 ml) was added, and the aqueous solution was extracted with     CH₂Cl₂. The combined organic extract was dried (MgSO₄), filtered,     and concentrated. Purification by silica gel chromatography (eluant:     5% EtOAc—CH₂Cl₂) gave 4.50 g (19.3 mmol, 64%) of the product 114A as     a beige foam. MS (M+2-tBu): m/e 178.

The following intermediates were synthesized by using a similar procedure:

Number Compound MS (M + 1) 114B

220M + 2-tBu 114C

180M + 2-tBu 114D

182M + 2-tBu 114E

182M + 2-tBu 114F

198M + 2-tBu 114G

200M + 2-tBu 114H

232M + 2-tBu 114I

235 114J

235

-   Step 3: Using the procedure of step 3 from Example 31, the following     intermediates were synthesized:

Number Compound MS (M + 1) 115A

134 115B

176 115C

136 115D

138 115E

138 115F

154 115G

156 115H

188 115I

135 115J

135

EXAMPLE 34

-   Step 1: Using the procedure of step 6 from Example 32, the following     intermediates were synthesized:

Number Compound MS (M + 1) 117A

178 117B

192

-   Step 2: To a solution of compound 117A (1.08 g, 6.09 mmol) dissolved     in EtOH (20 ml) was added 10% palladium on carbon catalyst (0.25 g)     and 1.73 M HCl in EtOH (10.6 ml, 18.3 mmol). The reaction mixture     was shaken on a Parr shaker under 50 psi of hydrogen pressure for     16 h. The catalyst was removed by filtration through celite and     washed with EtOH. The filtrate was concentrated to give 1.14 g (5.71     mmol, 93%) of the product 118A as a beige solid. MS (M−NH₂): m/e     147.

The following intermediate was synthesized by using a similar procedure:

Number Compound MS (M + 1) 118B

161M-NH₂

EXAMPLE 35

-   Step 1: To a solution of compound 119 (3.00 g, 14.9 mmol) dissolved     in dry DMF (60 ml) under a N₂ atmosphere was added NaH (60 wt % in     oil, 1.19 g, 29.8 mmol). The reaction mixture was stirred at RT for     30 min, then 2-chloropyrimidine (3.41 g, 29.8 mmol) was added. The     reaction mixture was heated at 80° C. for 16 h and then cooled to     RT. The solvent was evaporated. Water (75 ml) was added and the     aqueous solution was extracted with CH₂Cl₂. The combined organic     extract was dried (MgSO₄), filtered, and concentrated. Purification     by silica gel chromatography (eluant: 1%-4% MeOH—CH₂Cl₂) gave 2.49 g     (8.91 mmol, 60%) of the product 120A as a yellowish-orange solid. MS     (M+1): m/e 280.

The following intermediates were synthesized by using a similar procedure:

Number Compound MS (M + 1) 120B

280 120C

310

-   Step 2: Using the procedure of step 3 from Example 31, the following     intermediates were synthesized:

Number Compound MS (M + 1) 121A

180 121B

180 121C

210

EXAMPLE 36

-   Step 1: Trans-4-hydroxy-L-proline 122A (10.48 g, 80 mmol) was     refluxed in a 5-6 M solution of HCl in 2-propanol (200 ml) for 2 h.     The solvent was evaporated to give 16.33 g of the product 123A as a     white solid (97% yield). MS (M+1) 174. -   Step 2: 123A (16.33 g, 78.1 mmol) was suspended in dichloromethane     (460 ml). Et₃N (30 ml) and di-tert-butyl dicarbonate (20.33 g) were     added, and the mixture was stirred for 20 h at RT. The reaction     mixture was washed twice with equivolume 1 N HCl, once with     saturated NaHCO₃, and once with saturated NaCl. The organic solution     was dried (anhydrous Na₂SO₄), filtered, and concentrated to give the     product 124A as an amber oil (19.7 g, 92% yield). MS (M+1): m/e 274. -   Step 3: Using the procedure of step 5 from Example 32, compound 125A     was synthesized. MS (M+1): m/e 272. -   Step 4: A 5-gram vial of CeCl₃ from Aldrich was cracked open and     quickly added to a flame-dried, 125-ml, round-bottomed flask under     an Ar atmosphere. Anhydrous THF was added, and the mixture was     sonicated for 1 h and stirred an additional 1 h. The ketone 125A was     dissolved in dry THF (5 ml) and added to the CeCl₃/THF mixture and     stirred at RT for 1 h. In a separate round bottom flask, 2-pyrimidyl     tri-n-butylstannane was dissolved in dry THF (18 ml) under an Ar     atmosphere and cooled to −78° C. A 2.5M solution of n-butyllithium     in hexanes (4 ml) was added dropwise to the pyrimidyl stannane, and     the mixture turned thick and brown. After stirring for 1 h at −78°     C., this cold mixture was transferred via cannula to the ketone     125A/CeCl₃ mixture also cooled to −78° C. The resulting reaction     mixture was stirred at −78° C. for 3 h and then stirred at −50° C.     for 30 min. The mixture was again cooled to −78° C. and quenched     dropwise with 1 M citric acid (200 ml). The aqueous solution was     extracted with hexane and then Et₂O. The combined organic extract     was dried (MgSO₄), filtered, and concentrated. Purification by     silica gel chromatography gave 0.95 g of the product 126A (27%     yield). MS (M+1): m/e 352.

The following intermediates were synthesized by using a similar procedure:

Number Compound MS (M + 1) 126B

351 126C

280

-   Step 5: Using the procedure of step 3 from Example 31, the following     intermediates were synthesized:

Number Compound MS (M + 1) 127A

252 127B

251 127C

180

EXAMPLE 37

2,4,6-Trifluorobenzylamine 128 was prepared according to the literature procedure of A. Marfat et al, WO 9845268.

EXAMPLE 38

-   Step 1: Compound 129 (0.24 g, 0.5 mmol) was mixed with Et₃N (0.1 ml,     0.7 mmol) in dry THF (4 ml) and cooled to −78° C. Trimethylacetyl     chloride (0.08 ml, 0.6 mmol) was added, and the resulting mixture     was stirred at 0° C. for 20 min, then cooled to −78° C. again. In a     separate flask, oxazolidinone (0.07 g, 0.8 mmol) was dissolved in     dry THF (2 ml), cooled to −78° C., and 1.6 ml of a 1.6 M n-BuLi     solution in hexane was added. After stirring at −78° C. for 15 min,     the mixture was cannulated into the above mixed anhydride solution.     The resulting solution was then slowly warmed up to RT. The reaction     mixture was quenched with saturated NH₄Cl solution (2 ml). EtOAc     (50 ml) was added, and the organic solution was washed with 1 N HCl     solution, saturated NaHCO₃ solution, and brine. The organic solution     was dried (Na₂SO₄), filtered, and concentrated. Purification by     flash chromatography gave the product which was treated with 2 N HCl     in ether (50 ml) at RT overnight. The precipitate was collected by     filtration and dried in a vacuum oven at 50° C. overnight to give     0.15 gram of the product 130 as the HCl salt. MS (M+1): m/e 451.

EXAMPLE 39

-   Step 1: Compound 131 (1.15 g, 6 mmol) was mixed with Et₃N (0.9 ml,     6.4 mmol) in dry THF (20 ml), and cooled to −30° C. Trimethylacetyl     chloride (0.75 ml, 6 mmol) was added, and the resulting mixture was     stirred at −10° C. for 20 min. Pyrrolidine (0.85 ml, 10 mmol) was     added, and the resulting mixture was stirred at 0° C. for 30 min.     The reaction mixture was diluted with EtOAc (150 ml) and washed with     1 N HCl solution, saturated NaHCO₃ solution, and brine. The organic     solution was dried (MgSO₄), filtered, and concentrated to give the     product 132. -   Step 2: Using the procedure of step 3 from Example 31, intermediate     133 was synthesized. -   Step 3: To a solution of compound 129 (0.48 g, 1 mmol) dissolved in     DMF (4 ml) and CH₂Cl₂ (10 ml) at RT was added DIPEA (1 ml) and HATU     (0.6 g). After 5 min, compound 133 (HCl salt, 0.23 g, 1.3 mmol) was     added. The reaction mixture was stirred at RT for 30 min, then the     mixture was diluted with EtOAc (75 ml) and washed with 1 N HCl (50     ml), saturated NaHCO₃ (50 ml), and brine. The organic solution was     dried (Na₂SO₄), filtered, and concentrated. Purification by silica     gel chromatography gave the product 134. -   Step 4: Using the procedure of step 3 from Example 31, compound 135     was synthesized. MS (M+1): m/e 506.

EXAMPLE 40

-   Step 1: Using the procedure for step 3 from Example 39, compound 136     was synthesized. -   Step 2: To a solution of compound 136 (0.2 g, 0.35 mmol) dissolved     in dry CH₂Cl₂ (6 ml) was added DAST (0.1 ml, 0.7 mmol). The reaction     mixture was stirred at RT for 2 days, then quenched with saturated     NaHCO₃ (2 ml). The mixture was diluted with CH₂Cl₂ (75 ml) and     washed with water then 1 N HCl solution. The organic solution was     dried (MgSO₄), filtered, and concentrated. Purification by silica     gel chromatography gave the product 137. -   Step 3: Using the procedure of step 3 from Example 31, compound 138     was synthesized. MS (M+1): m/e 467

EXAMPLE 41

-   Step 1: Using the procedure of step 2 from Example 32, intermediate     140 was synthesized. MS (M+1): m/e 281. -   Step 2: Using the procedure of step 3 from Example 31, intermediate     141 was synthesized. MS (M+1): m/e 181. -   Step 3: To a solution of compound 141 (1.9 g, 8.2 mmol, a TFA salt)     and Et₃N (2.5 g, 24.6 mmol) in CH₂Cl₂ (32 ml) at 0° C. was added a     solution of 2-nitrophenylsulphonyl chloride (1.99 g, 9 mmol) in     CH₂Cl₂ (8 ml) over a period of 5 min. The reaction mixture was     stirred at 0° C. for 2 h, and then saturated NaHCO₃ solution was     added. The product was extracted with CH₂Cl₂, washed with brine     (1×70 ml), dried over Na₂SO₄, filtered, and concentrated to give an     oily residue. Purification by silica gel chromatography (Biotage     System, eluant: 40:1 CH₂Cl₂:MeOH) gave 3.11 g (8.4 mmol, 100%) of     the product 142 as an off white solid. MS (M+1): m/e 366. -   Step 4: The combined reaction mixture of compound 142 (730 mg, 2     mmol), K₂CO₃ (2.76 g, 20 mmol) and 1,2-dibromoethane (3.74 g, 20     mmol) in DMF (6 ml) was heated at 60° C. for 17 h, and then quenched     with water. The product was extracted with EtOAc (3×30 ml), and the     combined extract was washed with brine (3×60 ml), dried over Na₂SO₄,     filtered, and concentrated to give an oily residue. Purification by     preparative silica gel chromatography (eluant: EtOAc) gave 640 mg     (1.64 mmol, 82%) of the product 143 as an oil. MS (M+1): m/e 392. -   Step 5: To a solution of compound 143 (640 mg, 1.64 mmol) in CH₃CN     (13 ml) was added Cs₂CO₃ (1.6 g, 4.92 mmol) and PhSH (216 mg, 1.97     mmol). The reaction mixture was stirred at RT for 1 h, filtered, and     the solid was washed with CH₂Cl₂. The filtrate was concentrated to     give a yellow oil. Purification by silica gel chromatography     (Biotage System, eluant: 20:1 CH₂Cl₂:MeOH (with 4% NH₃) gave 210 mg     (1 mmol, 61%) of the product 144A as a colorless oil. MS (M+1): m/e     207.

The following intermediates were synthesized by using a similar procedure:

Number Compound MS 144B

242 144C

209

EXAMPLE 42

-   Step 1: To a solution of amine 145 (400 mg, 2 mmol) and Et₃N (202     mg, 2 mmol) in EtOH (10 ml) was added 1-bromo-2-fluoroethane (1.27     g, 10 mmol). The reaction mixture, charged in a pressurized tube,     was heated at 70° C. for 3 days. Mass spectroscopy was used to     monitor the reaction. The reaction mixture was concentrated, and     then water was added. The product was extracted with CH₂Cl₂ (3×40     ml), washed with brine (3×50 ml), dried over Na₂SO₄, filtered, and     concentrated to give of the product 146 (1.75 mmol, 87%) as an oil,     which was used without further purification. MS (M+1): m/e 247. -   Step 2: Using the procedure of step 3 from Example 31, the following     compounds were synthesized:

EXAMPLE 43

-   Step 1: To a solution of amide 148 (1.4 g, 7 mmol) in DMF (28 ml)     was added NaH (554 mg, 23.1 mmol, 60% in oil) in portions over a     period of 8 min. The reaction mixture was stirred at RT for 50 min,     then Etl (3.28 g, 21 mmol) was added over a period of 2 min. The     reaction mixture was stirred at RT for 15 h and then quenched with     ice-water. The product was extracted with EtOAc/CH₂Cl₂, washed with     brine (3×30 ml), dried over Na₂SO₄, filtered, and concentrated to     give an oily residue. Purification by silica gel chromatography     (Biotage System, eluant: 100:1 CH₂Cl₂:MeOH) gave 1.19 g of the     product 149 (5.2 mmol, 74%) as an oil. MS (M+1): m/e 229. -   Step 2: Using the procedure of step 3 from Example 31, the following     compounds were synthesized:

Number Compound MS 150A

129 150B

115 150C

143 150D

157 150E

191 150F

143 150G

155 150H

159 150I

247

EXAMPLE 44

-   Step 1: Using the procedure of step 1 from Example 2, intermediate     152 was synthesized. -   Step 2: Using the procedure of step 2 from Example 2, intermediate     153 was synthesized. -   Step 3: Using the procedure of step 2 from Example 3, intermediate     154 was synthesized. MS (M+1): m/e 174.

EXAMPLE 45

-   Step 1: 3,5-Dichloro-4-pyridinecarboxaldehyde (155, 0.44 g, 2.5     mmol) was mixed with allylamine (0.56 ml, 7.5 mmol), NaB(OAc)₃H (1.1     g, 5 mmol) and HOAc (0.15 ml) in 1,2-dichloroethane (10 ml). The     reaction mixture was stirred at RT for 20 h and then poured into     saturated NaHCO₃ solution (10 ml). The resulting-mixture was stirred     at RT for 30 min, and the product was extracted with ether (3×40     ml). The combined organic extract was dried (Na₂SO₄), filtered,     concentrated, and then purified by silica gel chromatography to give     0.46 g of the product 156 as an oil. MS (M+1): m/e 217. -   Step 2: Compound 156 (0.32 g, 1.47 mmol) was mixed with tetrakis     (triphenylphosphine) palladium (0) (20 mg) and     N,N-dimethylbarbituric acid (0.73 g, 4.4 mmol) in CH₂Cl₂ (35 ml).     The reaction mixture was heated at reflux for 15 h. CH₂Cl₂ (35 ml)     was added, and the organic solution was washed with saturated NaHCO₃     solution, dried (MgSO₄), filtered, and concentrated. Purification by     silica gel chromatography gave 0.25 g of the product 157 as an oil.     MS (M+1): m/e 177.

EXAMPLE 46

-   Step 1: To a solution of N-Boc-L-hydroxyproline ethyl ester 158 (7.0     g, 27 mmol) dissolved in CH₂Cl₂ (20 ml) was added 15% Dess-Martin     reagent in CH₂Cl₂ solution (112 g). The reaction mixture was stirred     at RT for 15 h. CH₂Cl₂ (100 ml) was added, and the organic solution     was washed with 6% NaHCO₃ solution, dried, filtered, and     concentrated. Purification by silica gel chromatography gave 6.5 g     of the product 159 as an oil. MS (M+1): m/e 258. -   Step 2: To a solution of compound 159 (1.1 g, 4.28 mmol) dissolved     in dry THF (25 ml) and cooled to −78° C. was added CH₃MgBr solution     (3.7 ml, 1.7 M in toluene/THF) dropwise. The reaction mixture was     stirred at −78° C. for 1 h, then slowly warmed up to −25° C. The     reaction was quenched by the addition of 5% HCl solution and then     warmed up to RT. The resulting mixture was extracted with EtOAc     (2×40 ml). The combined organic extract was dried (Na₂SO₄),     filtered, and concentrated. Purification by silica gel     chromatography gave 0.30 g of the product 160 as an oil. MS (M+1):     m/e 274. -   Step 3: Using the procedure of step 3 from Example 31, compound 161     was synthesized. MS (M+1): m/e 174.

EXAMPLE 47

Anhydrous CeCl₃ (1.85 g, 7.5 mmol) was suspended in THF (25 ml) under N₂ and stirred overnight at RT. EtMgBr (2.5 ml of 3.0 M in THF, 7.5 mmol) was added dropwise, and the reaction mixture was stirred at RT for 1 h. A solution of ketone 162 (463 mg, 2.5 mmol) dissolved in THF (5 ml) was added dropwise to the suspension, and the resulting mixture was stirred at RT for 2 h. The reaction mixture was treated with EtOAc (5 ml) for 30 min at 20° C. and 2 M HCl, respectively, followed by extraction with EtOAc (2×100 ml). The combined organic extract was washed with brine, dried (MgSO₄), filtered, and concentrated to give the crude intermediate. This intermediate was dissolved in minimal EtOAc and HCl (10 ml of 2 M in Et₂O) was added. The reaction mixture was stirred at RT overnight to give the product 163 as a precipitate. The precipitate was filtered, washed with EtOAc, and dried in vacuo to give the product 163 as brown solid (276 mg, 73%). (M+1): m/e 116.

EXAMPLE 48

-   Step 1: Compound 164 was synthesized according to the procedure of     Cowden, Organic Letters (2003), 5(23), 4497-4499. -   Step 2: Using the procedure of step 3 from Example 31, compound 166     was synthesized. MS (M+1): m/e 178.

EXAMPLE 49

-   Step 1: To a suspension of compound 164 (9.95 g, 107 mmol) in H₂O     (100 ml) and EtOH (10 ml) was added KOH (28 g, 500 mmol). The     resulting mixture was heated at reflux for 3.5 h and then cooled to     0° C. Concentrated HCl (50 ml) was added with caution. The resulting     mixture was concentrated, and the remaining aqueous layer was     extracted with Et₂O. The combined organic extract was washed with     brine, dried (MgSO₄), filtered, and concentrated to give the product     165 (12.6 g, 88%) as a yellow liquid. -   Step 2: To a solution of compound 165 (10.2 g, 89.2 mmol) dissolved     in t-BuOH (150 ml) was added Et₃N (14 ml, 100.6 mmol) and     diphenylphosphoryl azide (21 ml, 97.4 mmol). The reaction mixture     was heated at reflux overnight and then cooled to RT. The resulting     mixture was concentrated, diluted with EtOAc, washed with 1 N HCl     (150 ml), satd. NaHCO₃ (50 ml) and brine, dried (MgSO₄), filtered,     and concentrated. Purification by silica gel chromatography (eluant:     1:6 EtOAc:hexane) gave the product 166 (4.05 g, 25%) as a white     solid. -   Step 3: To a solution of compound 166 (3.0 g, 16.4 mmol) dissolved     in MeOH (200 ml), and cooled to −78° C. was bubbled ozone until the     light blue color persisted. Triphenyl phosphine (9.3 g, 35.5 mmol)     was added, and the reaction mixture was stirred at RT overnight. The     resulting mixture was concentrated. Purification by silica gel     chromatography (eluant: 1:3 EtOAc:hexane) gave the product 167 (2.95     g, 97%) as a white solid. MS (M+1): m/e 186. -   Step 4: To a solution of compound 167 (3.4 g, 18.4 mmol) dissolved     in THF (70 ml), and cooled to −78° C. was added L-selectride (1.0 M     in THF, 22.4 ml, 22.4 mmol) dropwise. The reaction mixture was     stirred at −78° C. for 2 h. Water was added, and the resulting     mixture was warmed up to RT. The solution was concentrated, and     water was added. The aqueous solution was extracted with EtOAc. The     combined organic extract was washed with brine, dried (MgSO₄),     filtered, and concentrated. Purification by silica gel     chromatography (eluant: 1:1 EtOAc:hexane) gave the product 168 (2.74     g, 80%) as a white foam. MS (M+1): m/e 188. -   Step 5: To a solution of compound 168 (1.0 g, 5.35 mmol) and     p-nitrobenzoic acid (0.98 g, 5.88 mmol) dissolved in THF (25 ml) was     added triphenylphosphine (2.1 g, 8.0 mmol) and DEAD (1.27 ml, 8.0     mmol) sequentially. The reaction mixture was stirred at RT     overnight. The resulting solution was concentrated. Purification by     silica gel chromatography (eluant: 1:5 EtOAc:hexane) gave the     product 169 (1.53 g, 85%) as a white solid. MS (M+1): m/e 237. -   Step 6: To a solution of compound 169 (1.53 g, 4.55 mmol) dissolved     in MeOH (30 ml) at 0° C. was added K₂CO₃ (0.24 g, 1.8 mmol). The     resulting suspension was stirred at 0° C. for 2 h and then     concentrated. Purification by silica gel chromatography (eluant: 1:5     EtOAc:hexane) gave the product 170 (0.71 g, 83%) as a white solid.     MS (M+Na⁺): m/e 210. -   Step 7: To a solution of compound 170 (0.85 g, 4.5 mmol) dissolved     in CH₂Cl₂ (40 ml) at 0° C. was added Et₃N (0.94 ml g, 6.7 mmol) and     mesyl chloride (0.45 ml, 5.8 mmol). The resulting solution was     stirred at 0° C. for 2 h. Water was added, and the resulting mixture     warmed up to RT. The aqueous layer was extracted with CH₂Cl₂. The     combined organic extract was washed with brine, dried (MgSO₄),     filtered, and concentrated to give the product 171 (1.0 g, 83%) as a     white solid. MS (M+Na⁺): m/e 288. -   Step 8: To a solution of compound 171 (1.0 g, 3.8 mmol) dissolved in     DMF (4 ml) was added NaN₃ (378 mg, 5.8 mmol). The reaction mixture     was heated at 85° C. overnight. The resulting solution was cooled to     RT, concentrated, and water was added. The aqueous layer was     extracted with EtOAc. The combined organic extract was washed with     brine, dried (MgSO₄), filtered, and concentrated to give the product     172 (0.78 g, 98%) as a white solid. MS (M+H⁺): m/e 213. -   Step 9: To a solution of compound 172 (0.78 g, 3.8 mmol) dissolved     in THF (30 ml) and H₂O (3 ml) was added triphenylphosphine (3.86 g,     14.7 mmol). The resulting solution was heated at reflux for 2 h,     cooled to RT, and then concentrated. Purification by silica gel     chromatography (eluant: 1:10 4% NH₃MeOH:CH₂Cl₂ then 1:2 4%     NH₃-MeOH:CH₂Cl₂) gave the product 173 (0.68 g, 100%) as a white     foam. MS (M+1): m/e 187.

EXAMPLE 50

-   Step 1: To compound 166 (1.0 g, 5.46 mmol was added 9-BBN (0.5 N in     THF, 16.4 ml, 8.2 mmol) dropwise. The reaction mixture was stirred     at RT overnight. The resulting mixture was cooled to 0° C., and     2-bromopyrimidine (1.3 g, 9.2 mmol), Pd(dppf)₂Cl₂ (446 mg, 0.55     mmol), K₂CO₃ (1.13 g, 8.19 mmol), DMF (6 ml), and water (0.44 ml)     were added. The reaction mixture was stirred at RT overnight. 0.5 N     NaOH (50 ml) was added and the mixture extracted with CH₂Cl₂. The     combined organic extract was washed with brine, dried (MgSO₄),     filtered, and concentrated. Purification by silica gel     chromatography (eluant: 1:3 EtOAc:hexane) gave a 4:1 mixture of 174A     and 174B (0.8 g, 56%) as a white solid. -   Step 2: Using the procedure of step 3 from Example 31, compounds     175A and 175B were synthesized.

EXAMPLE 51

-   Step 1: To a solution of compound 168 (374 mg, 2.0 mmol) dissolved     in DMF (5 ml) was added NaH (60% dispersion in mineral oil, 0.2 g, 5     mmol). The reaction mixture was stirred at RT for 30 min.     2-Bromopyrimidine (350 g, 2.2 mmol) was added, and the resulting     solution was stirred at RT for 4 h. The reaction mixture was     concentrated, and EtOAc and satd. NaHCO₃ (aq) were added. The     aqueous layer was separated and extracted with EtOAc. The combined     organic extract was washed with brine, dried (MgSO₄), filtered, and     concentrated. Purification by silica gel chromatography (eluant: 1:1     EtOAc:hexane) gave the product 176 (0.25 g, 47%) as a white solid.     MS (M+1): m/e 266. -   Step 2: Using the procedure of step 3 from Example 31, compound 177     was synthesized.

EXAMPLE 52

-   Step 1: To a solution of compound 168 (374 mg, 2.0 mmol) dissolved     in CH₃CN (8 ml) was added N,N′-disuccinimidyl carbonate (769 mg, 3.0     mmol) and Et₃N (0.84 ml, 6.0 mmol). The reaction mixture was heated     at 85° C. for 1 h. The resulting solution was concentrated, and     EtOAc and satd. NaHCO₃ (aq) were added. The aqueous layer was     separated and extracted with EtOAc. The combined organic extract was     washed with brine, dried (MgSO₄), filtered, and concentrated to give     the product 178 (0.25 g, 47%) as a white solid. MS (M+Na⁺): m/e 288. -   Step 2: To a solution of compound 178 (164 mg, 0.5 mmol) dissolved     in CH₃CN (8 ml) was added methylamine hydrochloride salt (68 mg, 1.0     mmol), Et₃N (0.45 ml, 3.3 mmol), and DMAP (2 mg). The reaction     mixture was stirred at RT overnight. EtOAc and satd. NaHCO₃ (aq)     were added. The aqueous layer was separated and extracted with     EtOAc. The combined organic extract was washed with brine, dried     (MgSO₄), filtered, and concentrated. Purification by silica gel     chromatography (eluant: 1:50 MeOH:CH₂Cl₂) gave the product 179 (60     mg, 49%) as a white solid. MS(M+H⁺−100): m/e 145. -   Step 3: Using the procedure of step 3 from Example 31, compound 177     was synthesized.

EXAMPLE 53

-   Step 1: To a suspension of compound 165 (6.22 g, 55.5 mmol) in DMF     (60 ml) was added Etl (26.0 g, 166 mmol) and Cs₂CO₃ (36 g, 111     mmol). The reaction mixture was stirred at RT overnight, then     diluted with Et₂O (200 ml) and washed with water (60 ml×3). The     aqueous layer was extracted with Et₂O. The combined organic extract     was washed with brine, dried (MgSO₄), filtered, and concentrated to     give the product 181 (7.2 g, 93%) as a light yellow oil. -   Step 2: Using the procedure for step 3 from Example 49, intermediate     182 was synthesized. -   Step 3: Using the procedure for step 1 from Example 45, intermediate     183 was synthesized. Purification of 183 by silica gel     chromatography (eluant: 1:20 EtOAc:hexane) gave the product 183A,     cis-isomer (2.07 g, 29%) as a colorless liquid; a mixture of cis and     trans isomer (183A and 183B) (2.54 g, 35%) as a colorless liquid.

MS (M+1): m/e 233.

-   Step 4: To a solution of compound 183A (2.0 g, 6.5 mmol) dissolved     in EtOH (25 ml) was added 4 N HCl in dioxane (0.25 ml) and Pd(OH)₂     catalyst (1.1 g). The reaction mixture was placed on a Parr shaker     under 50 psi of hydrogen pressure overnight. The resulting mixture     was filtered through celite. The filtrate was concentrated to give     the amine HCl salt (2.3 g). The amine HCl salt (1.04 g) was     suspended in CH₂Cl₂ (20 ml), and Et₃N (3.2 ml, 23.2 mmol) and Boc₂O     (0.76 g, 3.48 mmol) were added. The resulting mixture was stirred at     RT overnight, diluted with EtOAc and washed with 1 N HCl. The     aqueous layer was separated and extracted with EtOAc. The combined     organic extract was washed with brine, dried (MgSO₄), filtered, and     concentrated. Purification by silica gel chromatography (eluant: 1:6     EtOAc:hexane) gave the product 184A (0.34 g, 48%, two steps) as a     white solid. MS (M+Na⁺): m/e 266. -   Step 5: To a solution of compound 184A (0.15 g, 0.64 mmol) dissolved     in CH₂Cl₂ (6 ml), and cooled to −78° C. was added a solution of     DIBAL (1.0 M in CH₂Cl₂, 1.6 ml, 1.6 mmol) dropwise. The reaction     mixture was stirred at −78° C. to −40° C. for 2 h. The resulting     solution was warmed to RT, 10% potassium sodium tartrate solution     (4 ml) was added, and stirred for 30 min. The mixture was filtered,     and the filter cake was washed with CH₂Cl₂. The filtrate was washed     with brine, dried (MgSO₄), filtered, and concentrated. Purification     by silica gel chromatography (eluant: 1:6 EtOAc:hexane) gave the     product 185A (60 mg, 47%) as a colorless film. MS (M+Na⁺): m/e 222. -   Step 6: Using the procedure for step 6 from Example 32, intermediate     186A was synthesized. MS (M+1): m/e 215. -   Step 7: To a solution of compound 186A dissolved in THF (2 ml) was     added 1 M. LAH (0.3 ml, 0.3 mmol) dropwise under a N₂ atmosphere.     The reaction mixture was stirred at RT overnight. The resulting     solution was cooled to 0° C., and H₂O (50 μl), 15% NaOH (aq) (30     μl), and H₂O (0.5 ml) were added. The resulting slurry was stirred     at RT for 30 min and filtered through a pad of celite. The filtrate     was diluted with CH₂Cl₂ and washed with brine, dried (MgSO₄),     filtered, and concentrated to give the product 187A (34 mg, 97%) as     a white solid. MS (M+1): m/e 201.

EXAMPLE 54

-   Step 1: Compound 188 (8.80 g, 32 mmol) and ethyl     2-chloroacetoacetate (27.2 g, 23 ml, 160 mmol) were mixed together     and heated at 180° C. for 7 h. Excess ethyl 2-chloroacetoacetate was     removed by vacuum distillation. The residue was suspended in MeOH     (200 ml) and stirred at 60° C. for 40 min, then at RT overnight. The     solid was collected by vacuum filtration, washed with MeOH, and     dried under vacuum to give 8.5 g (74%) of the product 189 as a beige     solid. MS (M+1): m/e 381. -   Step 2: Using the procedure for step 1 from Example 2, intermediate     190 was synthesized. MS (M+1): m/e 459. -   Step 3: Compound 190 (0.20 g, 0.44 mmol) was suspended in 7 M NH₃ in     MeOH (10 ml) and heated at 55° C. for 16 h. The reaction mixture was     cooled to RT and concentrated. Purification by reverse phase     chromatography gave 35 mg (22%) of the title compound 191. MS (M+1):     m/e 367.

EXAMPLE 55

-   Step 1: Using the procedure for step 1 from Example 50, intermediate     192 was synthesized. MS (M+1): m/e 381. -   Step 2: Using the procedure for step 1 from Example 2, intermediate     193 was synthesized. -   Step 3: To a solution of compound 193 (2.0 g, 4 mmol) dissolved in     DMSO (20 ml) was added NaN₃ (0.29 g, 4.4 mmol). The reaction mixture     was stirred at RT for 24 h. Water was added and a precipitate     formed. The solid was collected by vacuum filtration, washed with     water, and dried under vacuum to give 1.7 g (92%) of the     product 194. MS (M+1): m/e 422. -   Step 4: To a solution of compound 194 (1.7 g, 4 mmol) dissolved in     toluene (30 ml) was added trimethylphosphine (1 M in toluene, 4.4     ml, 4.4 mmol). The reaction mixture was stirred at RT for 1 h and     then cooled to −20° C.     2-(tert-butoxycarbonyl-oxyimino)-2-phnylacetonitrile (BOC-ON) (1.18     g, 4.8 mmol) was added. The reaction mixture was warmed to RT and     stirred for 16 h. CH₂Cl₂ was added and the organic solution was     washed with water. The organic solution was dried (MgSO₄), filtered,     and concentrated. Purification by silica gel chromatography gave     1.26 g (64%) of the product 195. MS (M+1): m/e 496. -   Step 5: Using the procedure for step 1 from Example 32, intermediate     196 was synthesized. MS (M+1): m/e 482. -   Step 6 and Step 7: Using the procedure for step 2 from Example 32     and then step 3 from Example 31, the following compounds were     synthesized.

Number Compound MS (M + 1) 197A

541 197B

575

EXAMPLE 56

-   Step 1: Using the procedure for step 1 from Example 30, the compound     198 was synthesized. -   Step 2: To compound 198 (1.6 g, 5.6 mmol) dissolved in EtOH (50 ml)     was added ethyl 2-chloroacetoacetate (2.7 g, 2.3 ml, 16.8 mmol). The     reaction mixture was heated at 65° C. for 16 h and then cooled to     RT. The solid was collected by vacuum filtration and washed with     MeOH. Purification by silica gel chromatography gave the product     199. -   Step 3: Using the procedure for step 1 from Example 2, intermediate     200 was synthesized. MS (M+1): m/e 477. -   Step 4: Using the procedure for step 3 from Example 54, title     compound 201 was synthesized. MS (M+1): m/e 383.

EXAMPLE 57

-   Step 1: Using the procedure for step 1 from Example 54, compound 202     was synthesized. -   Step 2: Using the procedure for step 1 from Example 2, compound 203     was synthesized. -   Step 3: Using the procedure for step 3 from Example 55, compound 204     was synthesized. -   Step 4: Using the procedure for step 4 from Example 55, compound 205     was synthesized. -   Step 5: Using the procedure for step 1 from Example 32, intermediate     206 was synthesized. -   Step 6 and Step 7: Using the procedure for step 2 from Example 32     and then step 3 from Example 31, the following compounds were     synthesized.

Number Compound MS (M + 1) 207A

557 207B

591 207C

611 207D

397

EXAMPLE 58

-   Step 1: To a suspension of threonine-OMe-HCl (10.2 g, 0.06 mol) in     CH₂Cl₂ (200 ml) was added Hunig's base (14.1 g, 19 ml, 0.11 mol) and     the mixture cooled to 0° C. Compound 2 (15.0 g, 0.05 mol) dissolved     in CH₂Cl₂ (150 ml) was added dropwise via addition funnel. The     reaction mixture was stirred at 0° C. for 15 min, then at RT for 60     min. The solvent was evaporated and dilute HCl solution was added.     The solid was collected by vacuum filtration and washed with MeOH. A     second crop was collected by vacuum filtration of the filtrate. The     combined solid was dried under vacuum to give 19.3 g (100%) of the     product 208. -   Step 2: To a solution of compound 208 (7.7 g, 20 mmol) dissolved in     DMSO (50 ml) and toluene (50 ml) and cooled to 0° C. was added EDCI     (9.6 g, 50 mmol) and dichloroacetic acid (3.3 g, 2.1 ml, 25 mmol).     The reaction mixture was stirred at 0° C. for 5 min, then at RT for     45 min. Na₂S₂O₃ (7 g) dissolved in water (600 ml) and hexane     (300 ml) was added. The reaction mixture was stirred at RT for 15     min. The solid was collected by vacuum filtration and washed with     water, 1:1 water:MeOH, and then 1:1 ether:hexane. The filtrate was     filtered to give additional solid. The combined solid was dried     under vacuum to give 7.2 g (94%) of the product 209. -   Step 3: Using the procedure for step 1 from Example 30, the compound     210 was synthesized. -   Step 4: Using the procedure for step 1 from Example 2, the compound     211 was synthesized. -   Step 5: Using the procedure for step 2 from Example 2, the compound     212 was synthesized. -   Step 6: Using the procedure for step 3 from Example 2, the compound     213 was synthesized. -   Step 7 and Step 8: Using the procedures for step 1 and step 2 from     Example 3, the following compounds were synthesized.

Number Compound MS (M + 1) 214A

543 214B

577 214C

383

EXAMPLE 59

-   Step 1: To a solution of compound 215 (1.89 g, 10 mmol) in anhydrous     CH₂Cl₂ (25 ml) at −20° C. was added pyridine (790 mg, 10 mmol),     followed by the addition of cyanuric fluoride (3.6 ml, 40 mmol) over     a period of 5 min. After 2 h at −20° C., the reaction mixture was     quenched with ice-water and extracted with CH₂Cl₂. The combined     extract was washed with brine, dried over Na₂SO₄, filtered, and     concentrated to give 1.15 g (6 mmol, 60%) of the product 216 as a     colorless liquid. -   Step 2: To a solution of compound 217 (1.54 g, 3.98 mmol) and 216     (920 mg, 4.81 mmol) in anhydrous THF (16 ml) at −78° C. was added     KN(TMS)₂ (20 ml, 20 mmol) over a period of 5 min. After 1 h at −78°     C., the cold bath was removed and the reaction mixture was stirred     for another 30 min, quenched with water, and extracted with EtOAc.     The combined extract was washed with brine, dried over Na₂SO₄,     filtered, and concentrated to give an oily residue. Purification by     silica gel chromatography (Biotage System, eluant: 3:1 hexane:EtOAc)     gave 0.89 g (2.1 mmol, 54%) of the product 218 as a white powder. MS     (M+1): m/e 510. -   Step 3: Using the procedure for step 1 from Example 32, intermediate     219 was synthesized. MS (M+1): m/e 482. -   Step 4 and Step 5: Using the procedure for step 2 from Example 32     and then step 3 from Example 31, the following compounds were     synthesized:

Number Compound MS 220A

507 220B

558 220C

605 220D

572 220E

537

EXAMPLE 60

-   Step 1: To a solution of compound 221 (6.01 g, 42.9 mmol) dissolved     in DMF (50 ml) was added N-iodosuccinimide (10.27 g, 45.6 mmol). The     solution was heated at 40° C. overnight. The reaction was followed     by taking ¹H NMR of small amounts of the reaction mixture.     Additional N-iodosuccinimide (1.34 g, 5.96 mmol) was added, and the     resulting solution was stirred at RT for 2 days. The solution was     diluted with EtOAc (150 ml) and washed with 0.5 N Na₂S₂O₃ (50 ml×2).     The combined aqueous wash was extracted with EtOAc (100 ml×2). The     combined organic extract was dried (MgSO₄), filtered, and     concentrated. The residue was purified by silica gel chromatography     (eluant: 1:20 EtOAc:hexane) to give the product 222 (8.35 g, 73%) as     a light yellow liquid. MS (M+1): m/e 367. -   Step 2 and Step 3: To a solution of compound 222 (8.33 g, 31.3 mmol)     dissolved in CCl₄ (100 ml) was added NBS (11.1 g, 62.3 mmol) and     benzoylperoxide (1.3 g, 5.36 mmol). The reaction mixture was heated     at reflux for 16 h and then cooled to RT. CH₂Cl₂ was added (400 ml)     and the organic solution was washed with 0.5 N Na₂S₂O₃ (150 ml×2).     The aqueous washes were combined and extracted with CH₂Cl₂ (100     ml×3). The combined organic extract was washed with brine, dried     (MgSO₄), filtered, and concentrated. The residue was dissolved in     acetone (300 ml) and water (150 ml), and Ag₂CO₃ (10.3 g, 37.4 mmol)     was added. The reaction mixture was heated at reflux overnight and     then cooled to RT. The mixture was filtered through a pad of celite.     The filtrate was concentrated, and the remaining aqueous solution     was extracted with EtOAc. The combined organic extract was washed     with brine, dried (MgSO₄), filtered, and concentrated. The residue     was purified by silica gel chromatography (eluant: 1:3 EtOAc:hexane)     to give the product 224 (5.25 g, 60%) as a light yellow liquid. MS     (M+1): m/e 283. -   Step 4: To a solution of compound 224 (4.57 g, 16.2 mmol) dissolved     in CH₂Cl₂ (100 ml) was added Dess-Martin reagent (14 g, 33 mmol).     The reaction mixture was stirred at RT overnight. The resulting     solution was washed with 1 N NaOH (150 ml). The aqueous layer was     separated and extracted with CH₂Cl₂. The combined organic extract     was washed with brine, dried (MgSO₄), filtered, and concentrated.     The residue was purified by silica gel chromatography (eluant: 1:5     EtOAc:hexane) to give the product 225 (5.25 g, 60%) as a white     solid. MS (M+1): m/e 281. -   Step 5: To a solution of compound 226 (14 g, 51.7 mmol) dissolved in     quinoline (100 ml) was added copper (17 g, 268 mmol). The reaction     mixture was heated at 180° C. for 6 h and then cooled to RT. The     resulting mixture was filtered through a pad of celite and the     filter cake was washed with EtOAc. The filtrate was washed with 4 N     HCl (800 ml). The aqueous layer was separated and extracted with     EtOAc. The combined organic extract was washed with brine, dried     (MgSO₄), filtered, and concentrated. The residue was purified by     silica gel chromatography (eluant: 1:10 EtOAc:hexanes) to give the     product 227 (9.25 g, 79%) as a white solid. MS (M+1): m/e 228. -   Step 6: To a solution of compound 227 (9.1 g, 40.0 mmol) dissolved     in MeOH (200 ml) was added bromine (2.1 ml, 41.0 mmol). The reaction     mixture was heated at 40° C. for 2 h and then cooled to RT and     concentrated. The residue was purified by silica gel chromatography     (eluant: 1:6 EtOAc:hexane) to give the product 228 (12.1 g, 99%) as     a white solid. MS (M+1): m/e 306. -   Step 7: Pd₂(dba)₃ (1.69 g, 1.85 mmol) and 1.0 M PCy₃ in THF (3.87     ml, 3.87 mmol) were added to a 500 ml three-neck reaction flask     (evacuated and backfilled with N₂). Dioxane (200 ml) was added and     the mixture was evacuated and refilled with N₂ again. The resulting     mixture was stirred at RT for 30 min. Bromide 228 (5.91 g, 19.4     mmol), bis(pinocolo)diboron (6.88 g, 27.1 mmol), and KOAc (6.89 g,     70.0 mmol) were added sequentially. The reaction mixture was heated     at 85° C. overnight and then cooled to RT. The resulting mixture was     filtered through a pad of celite and the filter cake was washed with     EtOAc. The filtrate was washed with H₂O (100 ml). The aqueous layer     was separated and extracted with EtOAc. The combined organic extract     was washed with brine, dried (MgSO₄), filtered, and concentrated.     The residue was purified by silica gel chromatography (eluant: 1:15     EtOAc:hexane) to give the product 229 (5.35 g, 78%) as a white     solid. MS (M+1): m/e 354. -   Step 8: Boronic ester 229 (5.35 g, 15.15 mmol), 2-iodofuran 225     (4.27 g, 1.5.25 mmol), palladium acetate (172 mg, 0.77 mmol), S-Phos     (682 mg, 1.65 mmol), and K₃PO₄ (12.5 g, 54.3 mmol) were combined in     a 100 ml round bottom flask. The mixture was suspended in THF (100     ml), degassed, and refilled with N₂. Water (0.55 ml, 30 mmol) was     added. The resulting mixture was stirred at RT under a N₂ atmosphere     overnight. The reaction mixture was filtered through celite and the     filter cake was washed with EtOAc. The filtrate was concentrated,     and the residue was purified by silica gel chromatography (eluant     1:3 EtOAc:hexane) to give the product 230 (3.00 g, 46%) as a yellow     solid. MS (M+1): m/e 433. -   Step 9: To a solution of compound 230 (1.1 g, 2.90 mmol) dissolved     in CH₃CN (60 ml) and CH₂Cl₂ (15 ml) was added BocNH₂ (1.02 g, 8.71     mmol), Et₃SiH (1.4 ml, 8.76 mmol), and TFA (0.43 ml, 5.79 mmol)     sequentially. The reaction mixture was stirred at RT overnight. The     resulting solution was diluted with CH₂Cl₂ and washed with 1 N NaOH     (40 ml). The aqueous layer was separated and extracted with CH₂Cl₂.     The combined organic extract was washed with brine, dried (MgSO₄),     filtered, and concentrated. The residue was purified by silica gel     chromatography (eluant: 1:3 EtOAc:hexane) to give the product 231     (0.95 g, 68%) as a yellow solid. MS (M+1): m/e 481. -   Step 10: Using the procedure for step 1 from Example 32, compound     232 was synthesized. MS (M+1): m/e 467. -   Step 11 and Step 12: Using the procedure for step 2 from Example 32     and for step 3 from Example 31, the following compounds were     synthesized:

Number Compound MS 233A

492 233B

525 233C

496 233D

512 233E

506 233F

508 233G

520 233H

482

EXAMPLE 61

-   Step 1: Starting aldehyde 230 (1.21 g, 2.79 mmol),     t-(R)-butanesulfinylamide (400 mg, 3.30 mmol), and titanium ethoxide     (5.6 ml, 27 mmol) were mixed in dry THF (40 ml), degassed, and     refluxed under a N₂ atmosphere overnight. The reaction mixture was     cooled to RT and poured into brine (40 ml) with vigorous stirring.     The resulting mixture was filtered through celite. The filtrate was     extracted with EtOAc. The combined organic extract was washed with     brine, dried (MgSO₄), filtered, and concentrated. The residue was     purified by silica gel chromatography (Biotage, 40S+, eluant: 1:3     EtOAc:hexane) to give the product 234 (1.05 g, 76%) as a yellow     solid. MS (M+1): m/e 497. -   Step 2: To a solution of compound 234 (0.60 g, 1.2 mmol) dissolved     in dry THF (40 ml) under a N₂ atmosphere and cooled to −40° C. was     added a solution of MeMgBr (3 M in Et₂O, 0.5 ml, 1.5 mmol) dropwise.     The reaction mixture was stirred at −40° C. for 5 h and warmed up     overnight. The mixture was diluted with EtOAc, poured into saturated     NH₄Cl (aq) and filtered through celite. The aqueous layer was     separated and extracted with CH₂Cl₂. The combined organic extract     was washed with brine, dried (MgSO₄), filtered, and concentrated.     The residue was purified by silica gel chromatography (Biotage,     40S+, eluant: 1:1 EtOAc:hexane) to give the separated isomer 235A     (0.41 g, 66%) as a yellow solid MS (M+1): m/e 513, and isomer 235B     (0.10 g, 16%) as a yellow solid MS: (M+1): m/e 513.

Step 3: Using the procedure of step 1 from Example 32, the isomers 236A (from 235A) and 236B (from 235B) were synthesized. MS (M+1): m/e 485.

Step 4 and Step 5: Using the procedure of step 2 from Example 32 and then step 3 from Example 31, the compounds 237A (from 236A) and 237B (from 236B) were synthesized. MS (M+1): m/e 472.

EXAMPLE 62

Step 1: Solid t-BuOK (2.20 g, 20 mmol) was dissolved in dry THF (50 ml) and cooled to −78° C. Compound 239 (5.34 g, 20 mmol) dissolved in dry THF (20 ml) was added while the reaction mixture was maintained at −78° C. After stirring at −78° C. for 30 min, the solution was cannulated into a vigorously stirred solution of compound 238 (20 mmol) dissolved in dry THF (50 ml) and also cooled to −78° C. The reaction mixture was stirred at −78° C. for 30 min, then 3 N aqueous HCl solution (50 ml) was added, and the reaction mixture was stirred at RT for 1 h. The resulting mixture was concentrated, and the aqueous solution was washed with Et₂O (2×75 ml). The aqueous solution was concentrated by co-evaporation with toluene at temperature <40° C. The residue was dried under vacuum overnight, then suspended in MeOH (500 ml) and stirred at RT. The insoluble salt was removed by filtration. The filtrate was concentrated, and dried in a vacuum oven at 50° C. overnight to give the product 240 (6.6 g, 76%, HCl salt) as a solid. MS (M+1): m/e 397.

Step 2: To a solution of compound 241 (10 mmol) dissolved in dry THF (60 ml) and cooled to −78° C. was added compound 240 (4.3 g, 10 mmol) dissolved in dry DMF (30 ml) and then Et₃N (2.7 ml, 20 mmol). The reaction mixture was stirred at RT for 3 days. The resulting mixture was concentrated and the residue was dissolved in EtOAc/Et₂O. The organic solution was washed with 1 N HCl, 10% NaHCO₃, and brine, dried (Na₂SO₄), filtered, and concentrated. Purification by silica gel chromatography gave 242 (4.2 g, 65%) as pale solid. MS (M+1): m/e 650.

Step 3: Compound 242 (2.0 g, 3 mmol) was dissolved in dry p-xylene (60 ml) and 7 N NH₃/MeOH (2 ml) and TFA (2.2 ml) was added. The reaction mixture was heated at 150° C. for 2 h and then 0.5 N NH₃/dioxane (15 ml) and AcOH (2 ml) were added. The resulting mixture was heated at 160° C. with azeotropic removal of water overnight, cooled to RT, and concentrated. Purification by silica gel chromatography (20% EtOAc in CH₂Cl₂) gave the product 243 (0.51 g, 27%) as a light-yellow solid. MS (M+1): m/e 631.

Step 4: Compound 243 (0.46 g, 0.73 mmol) was dissolved in AcOH (20 ml) and concentrated HCl (10 ml) and was heated to reflux for 24 h. The resulting mixture was concentrated and water (50 ml) was added. The precipitate was collected by filtration, washed with water, and dried in a vacuum oven at 50° C. overnight, to give the product 244 (0.41 g, 93%). MS (M+1): m/e 603.

Step 5: To a solution of compound 244 (0.12 g, 0.2 mmol) dissolved in dry DMF (0.5 ml) and CH₂Cl₂ (3 ml) was added 2,4-difluorobenzylamine (0.05 ml, 0.4 mmol), DIPEA (0.07 ml, 0.4 mmol), and HATU (0.114 g, 0.3 mmol). The resulting mixture was stirred at RT overnight and then concentrated. The residue was dissolved in DMF (2 ml) and purified by Gilson reverse phase prep HPLC to give the product 245 (0.081 g, 56%). MS (M+1): m/e 728.

Step 6: Compound 245 (0.080 g, 0.11 mmol) was dissolved in Et₂NH (2 ml) and CH₃CN (2 ml) and stirred at RT for 30 min. The resulting mixture was concentrated, and the residue was purified by Gilson reverse phase prep HPLC. The product was treated with HCl in ether, then dried in a vacuum oven at 50° C. overnight to give the product 246 (0.052 g, 94%) as a di-HCl salt. MS (M+1): m/e 506.

EXAMPLE 63

Step 1: To a solution of compound 247 (38 g, 0.19 mol) dissolved in CH₂Cl₂ (450 ml) and cooled to 0° C. was added Et₃N (35 ml, 0.25 mol) and t-Boc anhydride (54 g, 0.25 mol). The reaction mixture was stirred at RT overnight. The resulting mixture was diluted with CH₂Cl₂, washed with 1 N HCl solution, dried (Na₂SO₄), filtered, and concentrated. Purification by silica gel chromatography gave the product 248 (47 g, 96%). MS (M+1): m/e 260.

Step 2: To a solution of compound 248 (1.55 g, 6 mmol) dissolved in dry THF (60 ml) and cooled to 0° C. was added triphenylphosphine (2.0 g, 7.8 mmol), diethyl azodicarboxylate (1.3 ml, 7.8 mmol) dropwise, and then diphenylphosphoryl azide (1.7 ml, 7.8 mmol). The reaction mixture was stirred at RT overnight, then diluted with ether. The organic solution was washed with saturated NaHCO₃ and brine, dried (Na₂SO₄), filtered, and concentrated. Purification by silica gel chromatography (eluant: 15-20% EtOAc in hexane) gave compound 249 (1.7 g, 100%). MS (M+1): m/e 285.

Step 3: To a solution of compound 249 (0.5 g, 1.76 mmol) dissolved in THF (40 ml) was added 10% Pd/C catalyst (0.25 g). The reaction mixture was stirred under H₂ (1 atm) at RT overnight. The resulting mixture was filtered, and the filtrate was concentrated to give the product 250 (0.45 g, 100%). MS (M+1): m/e 259.

Step 4 and 5: To a solution of compound 250 (0.13 g, 0.5 mmol) dissolved in CH₂Cl₂ (1 ml) was added DIPEA (0.2 ml) and cyclopropanecarbonyl chloride (0.053 ml, 0.5 mmol). The reaction mixture was stirred at RT for 2 h. The resulting mixture was diluted with EtOAc. The organic solution was washed with 1 N HCl, saturated NaHCO₃, and brine, dried, filtered, and concentrated. Purification by silica gel chromatography gave the product 251. Compound 251 was treated with 4 N HCl in dioxane at RT for 4 h. The resulting mixture was concentrated, and the residue was dried under vacuum for 2 days to give the product 252 as the HCl salt (0.1 g, 76%). MS (M+1): m/e 227.

Step 6: To a solution of compound 248 (3.1 g, 12 mmol) dissolved in dry THF (100 ml) and cooled to 0° C. was added triphenylphosphine (4.0 g, 15 mmol), DEAD (2.5 ml, 15 mmol) dropwise, and LiBr (5 g, 57 mmol). Within 2 min, all the LiBr dissolved. The resulting clear yellow solution was stirred at RT overnight. The reaction mixture was diluted with EtOAc, washed with water, dried (Na₂SO₄), filtered, and concentrated. Purification by silica gel chromatography gave the product 253 (2.15 g, 56%). MS (M+1): m/e 323.

Step 7: To a solution of compound 253 (2.1 g, 6.5 mmol) dissolved in DMSO (15 ml) was added NaN₃ (0.46 g, 7 mmol). The resulting mixture was stirred at RT for 2 days. Water was added to the mixture and product was extracted with ether (3×40 ml). The combined organic layer was washed with brine, dried (Na₂SO₄), filtered, and concentrated to give the product 254.

Step 8: Using the procedure of step 3, compound 255 was synthesized. MS (M+1): m/e 259.

Step 9: To a solution of compound 255 (0.26 g, 1 mmol) dissolved in DMF (2 ml) was added Et₃N (0.28 ml, 2 mmol) and 2-bromopyrimidine (0.16 g, 1 mmol). The reaction mixture was heated at 100° C. overnight then cooled to RT. The resulting mixture was diluted with DMSO (3 ml) and purification by reverse phase Gilson prep HPLC gave the product 256 (0.18 g, 54%). MS (M+1): m/e 337.

Step 10: Using the procedure of step 5, compound 257 was synthesized. MS (M+1): m/e 237.

EXAMPLE 64

Step 1: Compound 258 (4.14 g, 13.6 mmol), CuI (288 mg, 0.37 mmol), NaI (4.32 g, 28.8 mmol), and sym-dimethylethylenediamine (0.38 ml, 0.72 mmol) were suspended in toluene (12 ml). The reaction mixture was heated in a sealed tube at 125° C. for 48 h. The resulting mixture was cooled to RT and filtered through celite. The filtrate was concentrated, and the residue was purified by silica gel chromatography (eluant: 1:10 EtOAc:hexane) to give the product 259 (4.06 g, 85%) as a beige liquid. MS (M+1): m/e 354.

Step 2: Compound 259 (3.55 g, 10.0 mmol), pyrazole 260 (2.31 g, 15 mmol), trans-1,2-di(methylamine)cyclohexane (450 mg, 3.17 mmol), CuI (190 mg, 1.0 mmol), and K₂CO₃ (4.14 g, 30 mmol) were suspended in toluene (40 ml). The reaction mixture was heated in a sealed tube at 125° C. for 10 days. The resulting mixture was cooled to RT and filtered through celite. The filtrate was concentrated and the residue was purified by silica gel chromatography (eluant: 1:1 EtOAc:hexane) to give the starting compound 259 (2.1 g, 46%) and the product 261 (1.29 g, 45%) as a white solid. MS (M+1): m/e 380.

Steps 3, 4, and 5: Using procedures similar to that of step 1 from Example 2, step 3 from Example 55, and step 4 from Example 55, intermediate 262 was synthesized.

MS (M+1): m/e 495.

Step 6: Using a procedure similar to that of step 1 from Example 32, compound 263 was synthesized. MS (M+1): m/e 467.

Steps 7 and 8: Using procedures similar to that of step 2 from Example 32 and step 3 from Example 31, the following compounds were synthesized.

Number Compound MS 264A

492 264B

506

EXAMPLE 65

Using procedures from Examples 5 and 6, compound 266 was synthesized. MS (M+1): m/e 439.

The pharmacological activity of the compounds of the invention was measured by the following assays.

PDE4 Screening Assay

1. Human PDE4 Enzyme

The neutrophils were isolated from human blood using a standard procedure, then homogenized with a glass-glass homogenizer in a buffer containing 20 mM Tris/HCl (pH 8.0), protease inhibitor cocktail tablet (Cat. No. 1836145/Boehringer Mannheim), 2 mM EDTA, 1% Triton X-100 and 0.5% deoxycholate. After stirring for 2 h at 4° C., the samples were centrifuged at 100,000 g for 1 h. The supernatants were collected, filtered and applied to Mono Q column chromatography. The fractions containing the activity of hydrolyzing cAMP were determined and pooled as the enzymatic source of the PDE4 screening assay.

2. PDE4 Assay and Compound Screening

The PDE4 assays were performed using Phosphodiesterase [³H]cAMP SPA enzyme assay kits and its procedures (Cat. No. TRKQ 7090, Amersham). The assay procedures are described briefly as follows. The diluted PDE4 enzyme, 10× assay buffer and water were mixed at a ratio of 1:1:6 (10 μl/10 μl/60 μl). 80 μl aliquots of this mixture were added into the test wells of a 96-well Microlite plate (Cat. No. 7416, ThermoLabsystems). Enzyme dilution buffer, instead of the diluted enzyme, and water were added into the wells of negative control (background). 10 μl test compounds in 10% DMSO, standard inhibitor in 10% DMSO or 10% DMSO (for positive and negative controls) were added into the corresponding wells, respectively. After a 10 min incubation at RT, the reactions were initiated by addition of 10 μl pre-diluted [³H]cAMP into each well, then incubated at 30° C. for 30 min. The reactions were stopped by addition of 50 μl SPA beads into the test wells, then counted in a β-counter over 30 min ˜24 hr.

10× Assay Buffer: 500 mM Tris/HCl pH 7.5, 83 mM MgCl₂, 17 mM EGTA

[³H]cAMP: [3H] cAMP (40-60 Ci/mmol) is diluted at a 1:200 ratio with water. The final concentration is 0.005 μCi/μl

Yttrium SPA Beads: 500 mg of beads was reconstituted with 28 ml of water, stored at 4° C.

PDE10 and 11 Screening Assay

PDE10 (human recombinant PDE10A2, expressed in Sf9 insect cells by the baculovirus expression technique) was assayed using [³H]cGMP PDE SPA Assay kit (Amersham) at a final concentration of cGMP of 0.7 μM. PDE11 (human recombinant PDE11A3, expressed in Sf9 insect cells by the baculovirus expression technique) was assayed using [³H]cAMP PDE SPA Assay kit (Amersham) at a final concentration of cAMP of 0.0125 μM. Compounds were evaluated at 0.1-10,000 nM in 2% DMSO and 0.1% BSA from a stock solution of 4 mM in 100% DMSO. All assays were performed in duplicate, and each set of experiments was performed at least twice. Analysis of dose-response data and calculation of IC₅₀ values were performed using GraphPad Prism.

PBMC (Peripheral Blood Mononuclear Cell) Preparation and TNF Inhibition Assay

This protocol was modified from Prabhaker et al. (Int. J. Immunopharmac, Vol 16, No 10 pp 805-816, 1994. Smithkline Beecham Pharmaceuticals).

-   1. Human blood was collected from internal donors. The plasma was     separated from red blood cells by mixing with 6% dextran (4 ml for a     15-ml blood) and a 40 min-incubation at 37° C. -   2. 10 ml plasma was then layered on 9 ml Ficoll-paque (Cat. No.     17-1440-03, Amersham) in a centrifuge tube. -   3. After a centrifugation at 1500 rpm for 45 min, PBMC was removed     from the interface. -   4. PBMC was washed twice with PBS and counted. -   5. PBMC was suspended in RPMI medium containing 2.5%     heat-inactivated FCS (Hyclone laboratories Inc. Logan, Utah, USA),     Penicillin and streptomycin, and the cell volume was adjusted to     1×10⁶ cell/ml. -   6. 0.5 ml cells were transferred into each well of a 24 well plate. -   7. After one hour incubation at 37° C., the cells were pre-treated     for 1 h with 5 μl 10% DMSO (control) and 5 μl test compounds at     various concentrations (100 fold stock solutions in 10% DMSO). -   8. LPS was added to stimulate TNF production at a final     concentration of 100 ng/ml (E. coli 055:13S, SIGMA). -   9. The cells were stimulated for 14-16 h at 37° C. -   10. The supernatants were removed and transferred to new tubes. TNF     alpha levels were assayed by ELISA using Human TNF-α ELISA kit (Cat.     No. KHC3012, Biosource) and its procedures with an optimal dilution.     (1:10→1:100 dilution).     In vivo TNFα Assay

C57BI/6 mice were injected with 25 ug of LPS (LPS O55-B5, Sigma: L2880) by the intraperitoneal route. One hour prior to injection of the LPS, mice were treated orally with the PDE4 compounds at the selected doses. Ninety min after the LPS challenge, the mice were euthanized, and blood was collected through a heparinized syringe tip into Capijet T-MGA tubes. The blood was centrifuged for 10 min in a microcentrifuge at maximum speed (−13,000 rpm), and the serum was collected and analyzed for TNFα protein using an R&D ELISA kit.

Lipopolysaccharide (LPS) in vivo Assay

Male Sprague/Dawley rats (200-250 g) were purchased from Charles River Laboratories. Prior to use, the animals were permitted unrestricted access to food and water. Test compounds were delivered by gavage 5 hours prior to LPS-challenge. Compounds were suspended in a 0.4% methylcellulose vehicle with the same vehicle being given to control animals.

LPS-treatment: Animals were anethesized by inhalation of isoflurane, supplemented with oxygen (flow rate 1.0 ml/min). Once anesthetized, animals were placed supine and the trachea visualized using a small laryngoscope. Animals then received either 0.1 ml of saline or 0.1 ml of a 100 μg/ml lipopolysaccharide solution (LPS; E. coli) in saline by use of a Penn-Century Microspray needle (Penn-Century, Philadelphia, Pa.). Animals were allowed to recover on a heat pad, returned to housing and permitted access to food and water ad libitium. Sixteen hours after LPS-challenge, animals were anesthetized with an intra-peritoneal injection of the combination of ketamine/xylazine (10:1, 200 mg/kg ketamine, 20 mg/kg xylazine). After reaching anesthesia, animals were surgically prepared for bronchial lavage by inserting a tracheal cannula. Animals were lavaged with 2×2 ml of phosphate buffered saline, pH 7.2 (PBS). Routine recovery of BAL fluids did not significantly differ between animals with >80% of instilled volume recovered. Afterwards, animals were euthanized by surgically opening the thoracic cavity and cutting the diaphragm to assure lung collapse. Bronchial lavage (BAL) fluid was analyzed for cellular contents as described below.

BAL samples: Bronchial lavage (BAL) fluid was spun at 350×g for 10 min at 4° C. One ml of supernatant was removed and stored at −20° C. until analyzed for cytokine levels. Remaining fluid was aspirated and the cell pellet lysed for residual erythrocytes and resuspended in PBS, pH 7.2 containing 10 ug/ml of DNase I. Afterwards, the cell suspension was centrifuged at 350×g for 10 mins at 4° C., the supernatant aspirated and the cell pellet resuspended in 1 ml of PBS with 10 ug/ml DNase 1 and 5% heat inactivated fetal bovine serum. Cytospin slide preparations were made and stained with Hema3™ staining system (Fisher Scientific, Springfield N.J.). Differential cell counts were performed using standard histological parameters and at least 200 cells were enumerated. Total cell counts were performed using a Neubauer chamber.

Assay Procedure for Testing of Dermatitis in Dogs:

Five dogs are selected for each treatment group. Administration of experimental medications begins and continues through the end of the animal phase of the experiment. After three days, all dogs are sedated using medetomadine intravenously. An approximately 5 cm by 13 cm area is shaved on the lateral thorax of each dog. 1 cc of lidocaine is injected subcutaneously, and then two 8 mm punch biopsies are taken to act as Time 0 controls. Biopsy sites are closed with simple interrupted sutures of 3-0 Nylon suture.

Ten intradermal injections are given (five rows of two injections)—two injections are of phosphate buffered saline (PBS), and the remaining eight injections are of rabbit IgG antibody to dog IgE. Each injection is 0.05 ml. The total dose of anti-IgE per injection is 7 μg, as previously determined to be optimal. After injection, sites are observed and sampled. After injection and between all future samples, all dogs wear a protective garment (Quick Cover incision cover, Four Flags over Aspen) to prevent disturbance of the injection and/or biopsy sites.

The test compounds are compounds of formula I; the negative control is phosphate buffered saline (PBS); the positive control is commercially available prednisone tablets. Tablets are given orally by placement in the back of the mouth. Liquids are given by syringe to place the test article toward the back of the mouth. The dog's mouth may be held closed to ensure that all of the test article is swallowed. Plasma samples are analyzed for the concentration of test compound from the dogs treated with the active compounds. Samples from the negative control and prednisone treated dogs need not be analyzed.

Anti-IgE Site Observations: Sites of anti-IgE injection are examined and evaluated for erythema and wheal formation. At the 20 min observation time, the two PBS sites and the two 6 hr. biopsy sites are measured. At the other post-injection times, the two PBS sites and the corresponding biopsy sites are measured. If the size of the reaction is not consistent across sites in the same dog, then all sites that have not been previously biopsied will be measured. Wheals will be measured by calipers in two orthogonal dimensions as well as measured for thickness.

Collection of Skin Samples: Two 8 mm punch biopsies are taken of the sites injected with anti-IgE. One biopsy is placed in RNA isolation buffer and the other biopsy is bisected. One half goes into a standard 10% formalin solution for routine histopathological analysis and the other is deposited in Optimal Cutting Temperature Medium and quick frozen in liquid nitrogen, then maintained at −70° C. for immunohistochemical staining with Luna's stain for eosinophils, and Alcian Blue with Nuclear Fast Red counterstain for mast cells. Using manual or computerized morphometric analysis, the extent of infiltration by the following specific leukocytes is quantitated: CD 1a+c, IgE, CD3, 4+8, TCR alpha/beta and gamma/delta, TNF alpha, and TSLP. Cytokine analysis is to determine the presence of the following: TNF alpha, IL4, IL13, IL2, IFN gamma, and Thymic stromal lymphopoietin.

Allergic Brown Norway (BN) Rat Model:

Inbred male BN rats weighing 150 to 200 g were obtained from Charles River Laboratory (Wilmington, Mass.). Prior to use, the animals were allowed food and water ad libitum. The test compounds were administered 5 h prior to antigen challenge either by oral or inhalational route, as detailed in the “delivery of test compounds” section.

Sensitization and Antigen Bronchoprovocation

The animals were divided into two main groups viz. an alum group and an antigen group. In the antigen group, animals were sensitized by an intra-peritoneal (i.p.) injection of 1 ml alum-precipitated antigen containing 20 μg of ovalbumin (OVA, grade III; Sigma chemical Co., St Louis, Mo.) and 8 mg of Al(OH)₃ suspended in 0.9% saline vehicle. A booster injection of this alum-OVA mixture was given again 7 days later. Animals belonging to the alum group received injections containing alum only. Seven days after the second injection, animals were exposed to aerosolized antigen bronchoprovocation which was performed by placing the rats in an enclosed plexiglass chamber (21 liters) and exposing the rats to aerosolized OVA (1%) for 30 min. The aerosolized OVA was produced by an ultrasonic nebulizer (DeVilbiss, Somerset, Pa., USA; Model Ultra-Neb 99) at a flow rate of approximately 8 liters/min. Twenty four hours after aerosolized OVA challenge, the animals were euthanized with an overdose of pentobarbital sodium. The trachea was exteriorized and intubated, and the lungs were lavaged with two aliquots of 3 ml of physiological saline. The bronchoalveolar lavage fluid (BALF) thus collected was subjected to cell enumeration. Ten microliter of the BALF was utilized to manually enumerate the total white cells using a hemocytometer. One hundred microliter of BALF was used to prepare cytocentrifuge which was stained with Hema3™ staining system (Fisher Scientific, Springfield, N.J.) to identify and enumerate differential white blood cells such as eosinophils, neutrophils, mononuclear cells and epithelial cells. A total of 200 cells were enumerated from each cytocentrifuge. The ability of the compound to inhibit recruitment of inflammatory cells into the airways is reported.

Delivery of Test Compounds:

Oral administration: the compounds were dissolved in 0.4% methylcellulose and delivered to animals orally @3 ml/kg. An equivalent volume of 0.4% methylcellulose was given to both negative (alum group) and positive (antigen) control groups.

Intra-tracheal administration: the appropriate dose of the compound was mixed with lactose powder to achieve a final amount of 3 mg, which was delivered intra-tracheally to anesthetized animals. Animals were held in an upright position for 3-4 min and were allowed to recover from anesthesia before returning to their cages.

Using the procedures described above in the PDE 4, PDE10 and PDE11 screening assays, compounds of formula I were found to have IC₅₀ values for PDE4 in a range of 0.01 to 500 nM, with preferred compounds having a range of 0.01 to 100 nM, more preferably 0.01 to 10 nM, and most preferably 0.01 to 3 nM. Compounds of formula I are preferably selective PDE4 inhibitors compared to PDE10 and PDE11: preferably the IC₅₀ values for PDE10 and PDE 11 are 100 to 300 times the values for PDE4.

Representative compounds of formula I have the following IC₅₀ values for PDE4:

Compound IC₅₀ No. (nM) 13-106 0.14 26-42 0.07 26-92 0.01 26-177 3 26-241 0.2 26-293 0.5 26-417 1.4 26-444 0.03 38-3 1.8

For preparing pharmaceutical compositions from the compounds described by this invention, inert, pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, dispersible granules, capsules, cachets and suppositories. The powders and tablets may be comprised of from about 5 to about 70 percent active ingredient. Suitable solid carriers are known in the art, e.g. magnesium carbonate, magnesium stearate, talc, sugar, lactose. Tablets, powders, cachets and capsules can be used as solid dosage forms suitable for oral administration.

For preparing suppositories, a low melting wax such as a mixture of fatty acid glycerides or cocoa butter is first melted, and the active ingredient is dispersed homogeneously therein as by stirring. The molten homogeneous mixture is then poured into convenient sized molds, allowed to cool and thereby solidify.

Liquid form preparations include solutions, suspensions and emulsions. As an example may be mentioned water or water-propylene glycol solutions for parenteral injection.

Liquid form preparations may also include solutions for intranasal administration.

Aerosol preparations suitable for inhalation may include solutions and solids in powder form, which may be in combination with a pharmaceutically acceptable carrier, such as an inert compressed gas.

Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations for either oral or parenteral administration. Such liquid forms include solutions, suspensions and emulsions.

The compounds of the invention may also be deliverable transdermally. The transdermal compositions can take the form of creams, lotions, aerosols and/or emulsions and can be included in a transdermal patch of the matrix or reservoir type as are conventional in the art for this purpose.

Preferably the compound is administered orally or via inhalation.

Preferably, the pharmaceutical preparation is in unit dosage form. In such form, the preparation is subdivided into unit doses containing appropriate quantities of the active component, e.g., an effective amount to achieve the desired purpose.

The quantity of active compound of formula I in a unit dose of preparation may be varied or adjusted from about 0.1 mg to 1000 mg, more preferably from about 1 mg to 300 mg, according to the particular application.

The actual dosage employed may be varied depending upon the requirements of the patient and the severity of the condition being treated. Determination of the proper dosage for a particular situation is within the skill of the art. Generally, treatment is initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect under the circumstances is reached. For convenience, the total daily dosage may be divided and administered in portions during the day if desired.

The amount and frequency of administration of the compounds of the invention and the pharmaceutically acceptable salts thereof will be regulated according to the judgment of the attending clinician considering such factors as age, condition and size of the patient as well as severity of the symptoms being treated. A typical recommended dosage regimen for compounds of formula I is oral administration of from 10 mg to 2000 mg/day preferably 10 to 1000 mg/day, in two to four divided doses to provide relief from allergic and inflammatory diseases or the other disease or conditions listed above.

The doses and dosage regimen of the additional agents administered in the combinations of the invention will be determined by the attending clinician in view of the approved doses and dosage regimen in the literature, e.g., the package insert, taking into consideration the age, sex and condition of the patient and the severity of the disease.

While the present invention has been described in conjunction with the specific embodiments set forth above, many alternatives, modifications and variations thereof will be apparent to those of ordinary skill in the art. All such alternatives, modifications and variations are intended to fall within the spirit and scope of the present invention. 

1. A compound represented by the structural formula

or a pharmaceutically acceptable salt thereof, wherein

R is H or alkyl; X is O or S; R¹ is H, alkyl, cycloalkyl, cycloalkyl(C₁-C₄)alkyl-, —CH₂F, —CHF₂, —CF₃, —C(O)alkyl or —C(O)NR¹⁸R¹⁹; R³ and R⁴ are independently selected from the group consisting of H, alkyl, hydroxyalkyl and —C(O)Oalkyl; R⁵ and R⁶ are independently selected from the group consisting of H, alkyl, hydroxyalkyl, alkoxyalkyl, mercaptoalkyl, —CH₂F, —CHF₂, —CF₃, —C(O)OH, —C(O)Oalkyl and —C(O)NR⁴³R⁴⁴; t is 1 or 2; R⁷ is H, alkyl, alkenyl, hydroxyalkyl, cycloalkyl, alkoxyalkyl, aminoalkyl, (R¹⁷-phenyl)alkyl or —CH₂—C(O)—O-alkyl; R⁸ is H, alkyl, alkenyl, alkoxy, alkoxyalkyl, hydroxyalkyl, dihydroxyalkyl, alkyl-NR¹⁸R¹⁹, cyanoalkyl, haloalkyl, R²³-heteroaryl, R²³-heteroarylalkyl, R³⁶-heterocycloalkyl, (R³⁶-heterocycloalkyl)alkyl, R¹⁷-phenyl, (R¹⁷-phenyl)alkyl, R¹⁷-naphthyl, (R¹⁷-naphthyl)alkyl, R¹⁷-benzyloxy, -alkyl-C(O)—NR¹⁸R¹⁹, -alkyl-C(O)—N(R³⁰)—(R²³-heteroaryl), -alkyl-C(O)—(R¹⁷-phenyl), -alkyl-C(O)—(R³⁶-heterocycloalkyl); -alkyl-N(R³⁰)—C(O)Oalkyl, -alkyl-N(R³⁰)—C(O)—NR¹⁸R¹⁹, -alkyl-N(R³⁰)—C(O)alkyl, -alkyl-N(R³⁰)—C(O)-(fluoroalkyl), -alkyl-N(R³⁰)—C(O)—(R³⁹-cycloalkyl), -alkyl-N(R³⁰)—C(O)—(R¹⁷-phenyl), -alkyl-N(R³⁰)—C(O)—(R²³-heteroaryl), -alkyl-N(R³⁰)—C(O)-alkylene-(R²³-heteroaryl), -alkyl-NH—SO₂—NR¹⁸R¹⁹, -alkyl-N(R³⁰)—(R¹⁷-phenyl), -alkyl-N(R³⁰)—(R²³-heteroaryl), -alkyl-O—(R¹⁷-phenyl), -alkyl-O—(R²³-heteroaryl), -alkyl-N(R³⁰)—SO₂-alkyl, alkylthioalkyl-, alkyl-SO₂-alkyl-, (R³⁵-phenylalkyl)-S-alkyl-, (hydroxyalkyl)-S-alkyl-, (alkoxyalkyl)-S-alkyl-, -alkyl-CO₂-alkyl, R⁴⁵-hydroxyalkyl, dihydroxyalkyl substituted by R¹⁷-benzyloxy, dihydroxyalkyl substituted by R¹⁷-phenyl, alkoxyalkyl substituted by R¹⁷-phenyl, (R¹⁷-phenyl)alkyl substituted by —CO₂alkyl, (R¹⁷-phenyl)alkyl substituted by —C(O)N(R³⁰)₂, alkyl substituted by (R²³-heteroaryl) and —C(O)NR³⁷R³⁸, haloalkyl substituted by CO₂alkyl, R¹²-cycloalkyl, (R¹²-cycloalkyl)alkyl,

or R⁷ and R⁸ and the nitrogen to which they are attached together form a ring system selected from the group consisting of

comprises an R³⁵-substituted 5 or 6-membered heteroaryl group fused to the piperidinyl or pyrrolidinyl ring; p is 0 or 1; q is 0 or 1; the dotted line represents an optional double bond; R⁹ is H, halo, alkyl, cycloalkyl, —CH₂F, —CHF₂ or CF₃; R¹⁰, R¹¹, and R¹³ are independently selected from the group consisting of H and halo; R¹² is 1-3 substituents independently selected from the group consisting of H, alkyl, hydroxy, alkoxy, hydroxyalkyl, alkoxyalkyl, —C(O)Oalkyl, —(CH₂)_(n)—N(R³⁰)—C(O)-cycloalkyl, —(CH₂)_(n)—N(R³⁰)—C(O)alkyl, —(CH₂)_(n)—N(R³⁰)C(O)Oalkyl, —(CH₂)_(n)—N(R³⁰)—(R²³-heteroaryl), —(CH₂)_(n)—N(R³⁰)—C(O)—NR¹⁸R¹⁹, —(CH₂)_(n)—C(O)—NR¹⁸R¹⁹, R¹⁷-phenyl, R³⁵-heteroarylalkyl, R³⁵-heteroaryloxy, —C(O)-heterocycloalkyl, —O—C(O)-heterocycloalkyl, —O—C(O)—NR¹⁸R¹⁹, —NH—SO₂-alkyl, —NH—C(═NH)NH₂, and

or two R¹² substituents on the same carbon form ═O, ═NOR³⁰ or ═CH₂; R¹⁴ is 1 or 2 substituents independently selected from the group consisting of H, OH, halo, alkyl, alkoxy, hydroxyalkyl, alkoxyalkyl, —CF₃, CN, R¹⁷-phenyl, (R¹⁷-phenyl)alkyl, —NR¹⁸R¹⁹, alkyl-NR¹⁸R¹⁹, —(CH₂)_(n)—C(O)OH, —(CH₂)_(n)—C(O)Oalkyl, —(CH₂)_(n)—C(O)alkyl, —(CH₂)_(n)—C(O)(R³⁵-phenyl), —(CH₂)_(n)—C(O)(R²³-heteroaryl), —(CH₂)_(n)—C(O)NR¹⁸R¹⁹, (CH₂)_(n)—C(O)N(R³⁰)—(CH₂)_(n)—(R²³-heteroaryl), —(CH₂)_(n)—N(R³⁰)—C(O)alkyl, —(CH₂)_(n)—N(R³⁰)—C(O)-(fluoroalkyl), —(CH₂)_(n)—N(R³⁰)—C(O)-(cycloalkyl), —(CH₂)_(n)—N(R³⁰)—C(O)(R³⁵-phenyl), —(CH₂)_(n)—N(R³⁰)—C(O)(R²³-heteroaryl), —(CH₂)_(n)—N(R³⁰)C(O)NR¹⁸R¹⁹, —(CH₂)_(n)—N(R³⁰)—C(O)Oalkyl, —(CH₂)_(n)—N(R³⁰)cycloalkyl, —(CH₂)_(n)—N(R³⁰)(R¹⁷-phenyl), —(CH₂)_(n)—N(R³⁰)(R²³-heteroaryl), —(CH₂)_(n)—N(R¹⁸)SO₂alkyl, —(CH₂)_(n)—N(R²⁰)SO₂—(R¹⁷-phenyl), —(CH₂)_(n)—N(R³⁰)SO₂—CF₃, —CH₂S(O)₀₋₂(R³⁵-phenyl), —(CH₂)_(n)—OC(O)N(R³⁰)alkyl, R²³-heteroaryl, (R²³-heteroaryl)alkyl, (R²³-heteroaryl)oxy, (R²³-heteroaryl)amino, —CH(OH)—(R¹⁷-phenyl), —CH(OH)—(R²³-heteroaryl), —C(═NOR³⁰)—(R¹⁷-phenyl), —C(═NOR³⁰)—(R²³-heteroaryl), morpholinyl, thiomorpholinyl,

w is 0 or 1; or two R¹⁴ substituents and the carbon to which they are both attached form —C(═NOR³⁰)— or —C(O)—; each n is independently 0, 1, 2 or 3; R¹⁵ is H, alkyl, cycloalkyl, (cycloalkyl)alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl, —C(O)Oalkyl, —C(O)O(R³⁰-cycloalkyl), -alkyl-C(O)O-alkyl, —C(O)O-alkylene-(R³⁵-phenyl), R¹⁷-phenyl, (R¹⁷-phenyl)alkyl, —CH—(R¹⁷-phenyl)₂, R²³-heteroaryl, —(CH₂)_(n)—C(O)NR¹⁸R¹⁹, —SO₂-alkyl, —SO₂-cycloalkyl, —SO₂—CF₃, —SO₂—(R³⁵-phenyl), —SO₂—NR¹⁸R¹⁹, —C(O)alkyl, —C(O)-(fluoroalkyl), —C(O)—C(CH₃)(CF₃)₂, —C(O)—(R¹⁷-phenyl), —C(O)—(R²³-heteroaryl), —C(O)-hydroxyalkyl, —C(O)alkoxyalkyl, —C(O)—(R³⁹-cycloalkyl), —C(O)-alkylene-(R¹⁷-phenyl), —C(O)-alkylene-R²³-heteroaryl), —C(O)-alkylene-S—C(O)alkyl, —C(═S)—(R¹⁷-phenyl), hydroxyalkyl substituted by R¹⁷-phenyl, hydroxyalkyl substituted by R²³-heteroaryl, alkoxyalkyl substituted by R¹⁷-phenyl, alkoxyalkyl substituted by R²³-heteroaryl,

wherein z is 0, 1 or 2; R¹⁶ is 1 to 4 substituents independently selected from the group consisting of H, alkyl, R¹⁷-phenyl, (R¹⁷-phenyl)alkyl, (R²³-heteroaryl)alkyl, hydroxyalkyl, alkoxyalkyl and —(O)Oalkyl, or two R¹⁶ groups and the carbon to which they are both attached form —C(O)—; R¹⁷ is 1 to 3 substituents independently selected from the group consisting of H, halo, alkyl, cycloalkyl, —OH, hydroxyalkyl, alkoxy, alkoxyalkyl, —CN, —CF₃, —OCF₃, —OCHF₂, —OCH₂F, —C(O)OH, —C(O)Oalkyl, —C(O)O—(R³⁶-phenyl), —C(O)alkyl, —C(O)—(R³⁵-phenyl), —SOalkyl, —SO₂alkyl, —O₂—CF₃, alkylthio, —NR⁴³R⁴⁴, -alkyl-NR⁴³R⁴⁴, R³⁵-phenyl, R³⁵-phenoxy, R³⁵-heteroaryl, R³⁵-heteroaryloxy, R³⁶-heterocycloalkyl, —C(O)—(R³⁶-heterocycloalkyl), hydroxyalkyl-NH—, —C(O)N(R³⁰)₂, —N(R⁴³)—(R³⁵-cycloalkyl) and —C(═NOR³⁰); or two R¹⁷ substituents on adjacent carbon atoms together form —O—CH₂—O—, —O—(CH₂)₂—O—, —(CH₂)₂—O— or —O—CH₂—O—CH₂—; R¹⁸ and R¹⁹ are independently selected from the group consisting of H, alkyl, hydroxyalkyl, alkoxyalkyl, haloalkyl, R¹⁷-phenyl, (R¹⁷-phenyl)alkyl, naphthyl and cycloalkyl; R²⁰ is H, alkyl, or cycloalkyl; R²² is 1 to 4 substituents independently selected from the group consisting of H, alkyl, hydroxy, alkoxy, halo, —CF₃, —NH₂ and R³⁵-phenyl; R²³ is 1 to 4 substituents independently selected from the group consisting of H, alkyl, hydroxy, alkoxy, halo, —CF₃, —NR¹⁸R¹⁹, —CN, —C(O)Oalkyl, —SO₂-alkyl, —NHSO₂-alkyl, R³⁵-phenyl, R³⁵-heteroaryl, morpholinyl, and —(CH₂)_(n)—C(O)—N(R³⁰)₂; R²⁴ is H, OH or alkoxy; or when the optional double bond is present, R²⁴ and the adjacent carbon atom form the double bond; R²⁵ is H or R³⁵-phenyl; R²⁷ is 1 to 3 substituents independently selected from the group consisting of H, halo, OH, alkyl, alkoxy, hydroxyalkyl, alkoxyalkyl, haloalkyl, —CN, —C(O)OH, —C(O)Oalkyl, —C(O)N(R³⁰)(R¹⁸), —C(O)—(R³⁶-hetercycloalkyl), R¹⁷-phenyl, (R¹⁷-phenyl)-alkyl, R²³-heteroaryl, (R²³-heteroaryl)alkyl, (R²³-heteroaryl)oxy, (R²³-heteroaryl)amino NR¹⁸R¹⁹, NR¹⁸R¹⁹-alkyl, —(CH₂)_(n)—N(R³⁰)—C(O)alkyl, —(CH₂)_(n)—N(R³⁰)—C(O)-(fluoroalkyl), —(CH₂)_(n)—N(R³⁰)—C(O)alkoxyalkyl, —(CH₂)_(n)—N(R³⁰)—C(O)(cycloalkyl), —(CH₂)_(n)—N(R³⁰)—(R²³-heteroaryl), —(CH₂)_(n)—N(R³⁰)—C(O)—(R²³-heteroaryl), —(CH₂)_(n)—N(R³⁰)—C(O)O-alkyl, —(CH₂)_(n)—N(R³⁰)—C(O)O—(CF₃alkyl), —(CH₂)_(n)—N(R³⁰)—C(O)O—(R³⁹-cycloalkyl), —(CH₂)_(n)—N(R³⁰)—C(O)O-alkylene-cycloalkyl, —(CH₂)_(n)—N(R³⁰)—C(O)—N(R³⁰)(R²⁰), —(CH₂)_(n)—N(R³⁰)—SO₂-alkyl, —(CH₂)_(n)—N(R³⁰)—SO₂—CF₃, —(CH₂)_(n)—N(R³⁰)—SO₂—N(R³⁰)₂ and

or two R²⁷ groups and the carbon to which they are both attached form —C(═NOR³⁰)— or —C(O)—; R²⁸ is H, alkyl, R³⁵-benzyl or -alkyl-C(O)O-alkyl; R²⁹ is alkyl, haloalkyl, —C(O)Oalkyl, —C(O)alkyl, —C(O)CF₃, —C(O)—(R¹²-cycloalkyl), —C(O)—(R¹⁷-phenyl), —C(O)—(R²³-heteroaryl), —C(O)—(R³⁶-hetercycloalkyl), —SO₂-alkyl, —SO₂—(R³⁵-phenyl), —C(O)NR¹⁸R¹⁹, R³⁵-phenyl, (R³⁵-phenyl)alkyl or R²³-heteroaryl; R³⁰ is independently selected from the group consisting of H, alkyl, R³⁵-benzyl and R³⁵-phenyl; R³¹ is H, alkyl, R³⁶-benzyl or phenoxyalkyl; R³³ is H, OH or alkoxy; R³⁴ is H, alkyl, hydroxyalkyl, alkoxyalkyl or —C(O)Oalkyl; R³⁵ is 1 to 3 substituents independently selected from the group consisting of H, halo, alkyl, OH, —CF₃, alkoxy, —CO₂alkyl and —N(R⁴³)(R⁴⁴); R³⁶ is 1 or 2 substituents independently selected from the group consisting of H, alkyl, R¹⁷-phenyl, —OH, hydroxyalkyl, alkoxyalkyl, —C(O)Oalkyl and —NR¹⁸R¹⁹; or two R³⁶ groups and the carbon to which they are both attached form —C(═NOR³⁰)— or —C(O)—; R³⁷ and R³⁸ are independently selected from the group consisting of H and alkyl, or R³⁷ and R³⁸ together are —(CH₂)₃— or —(CH₂)₄—, and together with the nitrogen to which they are attached, form a ring; R³⁹ is H, OH, alkyl, alkoxy, or CF₃; R⁴⁰ is —OR³⁰ or —NHC(O)alkyl; R⁴¹ is H or —SO₂alkyl; R⁴² is —(CH₂)_(n)—(R³⁵-phenyl), —(CH₂)_(n)—(R²³-heteroaryl), —C(O)Oalkyl or —C(O)alkyl; R⁴³ and R⁴⁴ are independently selected from the group consisting of H and alkyl; and R⁴⁵ is 1 or 2 substituents independently selected from the group consisting of halo, alkoxyalkyl, —CO₂alkyl, R¹⁷-phenyl, R²³-heteroaryl and cycloalkyl.
 2. A compound of claim 1 wherein X is O.
 3. A compound of claim 2 wherein


4. A compound of claim 3 wherein R¹⁰, R¹¹ and R¹³ are each H; R¹ is H, alkyl, cycloalkyl or —CF₃; and R⁹ is H, alkyl or —CF₃.
 5. A compound of claim 4 wherein R¹⁰, R¹¹ and R¹³ are each H, R¹ is alkyl, and R⁹ is —CF₃.
 6. A compound of claim 5 wherein t is 1, R⁵ is H, R⁶ is H, alkyl or hydroxyalkyl, and R³ and R⁴ are each H or alkyl.
 7. A compound of claim 6 wherein R⁷ is H, alkyl, cycloalkyl, hydroxyalkyl or alkoxyalkyl, and R⁸ is R¹²-cycloalkyl, (R¹²-cycloalkyl)alkyl, R⁴⁵-hydroxyalkyl, R¹⁷-phenyl, (R¹⁷-phenyl)alkyl, R²³-heteroaryl, (R²³-heteroaryl)alkyl, -alkyl-N(R³⁰)—C(O)—NR¹⁸R¹⁹, -alkyl-N(R³⁰)—C(O)alkyl, -alkyl-N(R³⁰)—C(O)—(R¹⁷-phenyl), -alkyl-N(R³⁰)—C(O)—(R²³-heteroaryl), -alkyl-N(R³⁰)—(R²³-heteroaryl),


8. A compound of claim 7 wherein R⁸ is R¹²-cycloalkyl, R⁴⁵-hydroxyalkyl, (R¹⁷-phenyl)alkyl, R²³-heteroaryl, (R²³-heteroaryl)alkyl, -alkyl-N(R³⁰)—(R²³-heteroaryl), -alkyl-N(R³⁰)—C(O)alkyl,

wherein p is 0, or

wherein p is
 1. 9. A compound of claim 8 wherein R¹² is OH, —(CH₂)_(n)—N(R³⁰)—C(O)-cycloalkyl Or —(CH₂)_(n)—N(R³⁰)—(R²³-heteroaryl), R⁴⁵ is R¹⁷-phenyl, or R²⁹ is heteroaryl, —C(O)alkyl or —C(O)cycloalkyl.
 10. A compound of claim 6 wherein R⁷ and R⁸ and the nitrogen to which they are attached form


11. A compound of claim 10 wherein R⁷ and R⁸ form

the optional double bond is not present, and R¹⁴ is selected from the group consisting of H, OH, alkoxy, —(CH₂)_(n)—N(R³⁰)(R²³-heteroaryl), R²³-heteroaryl and (R²³-heteroaryl)-alkyl; or wherein R⁷ and R⁸ form

R¹⁵ is selected from the group consisting of alkyl, R¹⁷-phenyl, R²³-heteroaryl, —C(O)alkyl, —C(O)(fluoroalkyl), —C(O)—(R²³-heteroaryl), —C(O)-alkoxyalkyl, —C(O)—(R³⁸-cycloalkyl), —SO₂-alkyl, —SO₂—NR¹⁸R¹⁹ and

and R¹⁶ is selected from the group consisting of H and alkyl, or two R¹⁶ groups and the carbon to which they are attached form —C(O)—.
 12. A compound of claim 10 wherein R⁷ and R⁸ form

q is 1, and R²⁷ is 1, 2 or 3 substituents independently selected from the group consisting of H, OH, alkyl, alkoxy, alkoxyalkyl, R¹⁷-phenyl, —C(O)OH, —C(O)Oalkyl, R²³-heteroaryl, (R²³-heteroaryl)amino and —(CH₂)_(n)—N(R³⁰)—C(O)(cycloalkyl) wherein n is
 0. 13. A compound of claim 10 wherein R⁷ and R⁸ form

wherein p is 0, R³⁴ is H, and R³⁵ is 1 or 2 substituents independently selected from the group consisting of H, OH, halo and alkyl.
 14. A compound of claim 10 wherein R⁷ and R⁸ form

wherein p is 0, B is a pyrazolyl or thiazolyl ring, and R³⁵ is 1 or 2 substituents independently selected from the group consisting of H and alkyl.
 15. A compound of claim 7 wherein R⁷ is H or alkyl and R⁸ is (R¹⁷-phenyl)alkyl, R⁴⁵-hydroxyalkyl or -alkyl-N(R³⁰)—(R²³-heteroaryl), wherein R⁴⁵ is R¹⁷-phenyl; heteroaryl is pyridinyl, pyrimidinyl, pyrazinyl, indolyl, benzothienyl or benzofuranyl; R¹⁷ is 1 to 3 substituents independently selected from the group consisting of halogen, OH, alkoxy and alkyl; and R²³ is 1 or 2 substituents independently selected from the group consisting of H, alkyl, alkoxy and halogen.
 16. A compound of claim 1 selected from the group consisting of


17. The compound of claim 1 having the formula


18. The compound of claim 1 having the formula


19. The compound of claim 1 having the formula


20. The compound of claim 1 having the formula


21. The compound of claim 1 having the formula


22. The compound of claim 1 having the formula


23. The compound of claim 1 having the formula


24. The compound of claim 1 having the formula


25. The compound of claim 1 having the formula


26. A pharmaceutical composition comprising an effective amount of a compound of claim 1 and a pharmaceutically acceptable carrier.
 27. A method of treating a PDE4 mediated disease, where the disease is selected from COPD asthma, depression or multiple sclerosis comprising administering an effective amount of a compound of claim 1 to a patient in need of such treatment.
 28. The method of claim 27 for treating dermatitis in dogs.
 29. The method of claim 27 further comprising administering to said patient the compound of formula I in combination with at least one other medicament selected from the group consisting of disease modifying antirheumatic drugs, nonsteroidal anitinflammatory drugs, COX-2 selective inhibitors, COX-1 inhibitors, Immunosuppressives, steroids, biological response modifiers and other anti-inflammatory agents or therapeutics useful for the treatment of PDE4 mediated diseases. 